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J D Ostrow

Publications and source records attributed to J D Ostrow.

At least 55 records · Page 3Linked to original sources
PubMed1990

Unconjugated bilirubin and cholesterol gallstone formation.

Cholesterol gallstones usually have small amounts of pigment at their centers and often have diffuse pigmentation or pigmented layers alternating with cholesterol layers and/or pigmented rims associated with calcium carbonate (eggshell calcification). The pigments are primarily monomeric calcium salts of unconjugated bilirubin anions and/or an insoluble, black, network polymer of tetrapyrroles. Bilirubin presumably can precipitate only if bile is supersaturated with calcium bilirubinates. Among various in vitro model systems, the aqueous solubilities and pK'a values for unconjugated bilirubin differ greatly. It is therefore not known whether normal bile is saturated with unconjugated bilirubin. However, all systems indicate that unconjugated bilirubin is solubilized by binding to bile salt monomers and oligomers, as well as micelles; marked metastable supersaturation of unconjugated bilirubin can occur in the presence of bile salt micelles, and both pK'a values of unconjugated bilirubin are greater than 6.0, probably because of internal hydrogen-bonding of the--COOH groups. Lecithin decreases equilibrium solubilization of unconjugated bilirubin crystals but enhances metastable supersaturation of unconjugated bilirubin. Calcium ions form insoluble salts with unconjugated bilirubin monoanions and dianions but soluble complexes with bilirubin conjugates. The solubility products of the calcium bilirubinate salts suggest that normal hepatic bile is not saturated with CaB or Ca(HB)2 but that gallbladder bile may be supersaturated with Ca(HB)2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Binding of calcium by organic anions, determined by perturbation of the equilibrium solubility of [14C]calcium oxalate.

Confirmation is needed of the reported binding of calcium ions (Ca2+) by bile salts, which is believed to decrease the activity of free calcium ions [Ca2+] available for precipitation of insoluble calcium salts of organic anions in pigment gallstones. We report a new method to determine the association constants (K'f) of calcium for organic anions, from the perturbation by the added anion of the equilibrium solubilization of calcium [14C]oxalate monohydrate crystals (CaOx*). CaOx* crystals were prepared by stepwise conversion of [14C]oxalic acid to its K+ and Ca2+ salts. Structure and purity were confirmed by X-ray diffraction of the crystals. CaOx* was incubated (37 degrees C, under N2) in 0.15 M NaCl in CO2-free deionized H2O at pH 6.3. Dissolution of CaOx*, estimated by radioassay of the 0.22-micron Millipore filtrate, attained equilibrium at 3 days, with K'sp = [Ca2+] * [Ox=] = 2.34 X 10(-8) M2, calculated using known affinity constants for the soluble complexes of NaOx- (K'NaOx = 3.215 M-1) and CaOx (K'CaOx = 195.0 M-1). Keeping total [Na+] = 0.15 M, we added anions that formed soluble complexes with Ca2+. This decreased free [Ca2+], causing more CaOx* to dissolve in amounts related to the concentration of added anion and its K'f for Ca2+. With this method, K'f values for citrate, malonate and malate were similar to the values we determined with the Ca2+ ion electrode, and to published values obtained with the Ca2+ ion electrode and other methods. The sensitivity of the CaOx method permits determination of K'f values with small quantities and low concentrations of the anions and calcium.

Calcium↗
PubMed1989

Isolation of an acidic protein from cholesterol gallstones, which inhibits the precipitation of calcium carbonate in vitro.

In seeking to identify nucleating/antinucleating proteins involved in the pathogenesis of cholesterol gallstones, a major acidic protein was isolated from each of 13 samples of cholesterol gallstones. After the stones were extracted with methyl t-butyl ether to remove cholesterol, and methanol to remove bile salts and other lipids, they were demineralized with EDTA. The extracts were desalted with Sephadex-G25, and the proteins separated by PAGE. A protein was isolated, of molecular weight below 10 kD, which included firmly-bound diazo-positive yellow pigments and contained 24% acidic, but only 7% basic amino acid residues. The presence of N-acetyl glucosamine suggested that this was a glycoprotein. This protein at concentrations as low as 2 micrograms/ml, but neither human serum albumin nor its complex with bilirubin, inhibited calcium carbonate precipitation from a supersaturated solution in vitro. This protein could be precipitated from 0.15 M NaCl solution by the addition of 0.5 M calcium chloride. Considering that cholesterol gallstones contain calcium and pigment at their centers, and that small acidic proteins are important regulators in other biomineralization systems, this protein seems likely to play a role in the pathogenesis of cholesterol gallstones.

Amino Acids↗
PubMed1988

Interactions of unconjugated bilirubin with bile salts.

The rate of peroxidation of unbound, unconjugated bilirubin (UCB) was used to assess the interactions of UCB with four taurine-conjugated bile salts at pH 8.2, 37 degrees C, and an ionic strength of 0.15. Each of the four structurally different bile salts markedly decreased the rate of peroxidation of UCB in the presence of horseradish peroxidase (HRP); 30% of UCB was bound even at low, premicellar bile salt concentrations (1 mM). At high bile salt concentrations (75 mM), taurocholate (TC) and tauro-3 alpha,7 alpha-dihydroxy-12-oxo-5 beta-cholan-24-oate (T12-OXO) exhibited the highest degree of inhibition of UCB peroxidation; only 0.6% and 1.1% of UCB were unbound, respectively. Taurochenodeoxycholate (TCDC) yielded somewhat less inhibition with 2.0% of UCB unbound. Taurodehydrocholate (TDHC), a bile salt that does not form micelles but does form dimers, was comparable to TC and T12-OXO with unbound UCB of 1.0%. With TC and T12-OXO, apparent affinity for UCB was at least four times greater above the published critical micellar concentration (CMC) than in the premicellar range. TCDC was only studied above its CMC value and only one region of UCB binding was noted. Interaction of UCB with TDHC was similar to premicellar interactions with TC and T12-OXO below 25 mM, but increased to values intermediate between monomer and micelle above 40 mM TDHC, compatible with formation of TDHC dimers above 20 mM. These data show that there are differences in the ability of bile salts to bind UCB. Thus, alterations in bile salt profile in bile might lead to higher concentrations of free UCB in bile predisposing to pigment gallstones.

Bile↗
PubMed1988

Molecular and micellar associations in the pH-dependent stable and metastable dissolution of unconjugated bilirubin by bile salts.

Unconjugated bilirubin (UCB) is almost insoluble in water at neutral pH, but appears in normal human gallbladder bile at concentrations up to 35 microM. We therefore determined whether conjugated bile salts could increase the dissolved concentration [( Bt]) of UCB over the pH range 3.0-11.0. Using crystalline UCB, [Bt] was higher with less ordered crystals, with increasing pH and bile salt concentration, and with taurocholate (TC) micelles compared to taurodehydrocholate (TDHC) dimers. Plots of [Bt] verus pH from pH 3.0-9.3 fit the equation, [Bt] = A(1 + K'1/[H]+ + K'1.K'2/[H+]2), where A = [Bt] at pH less than 4.0, and K'1 and K'2 are the two apparent ionization constants of UCB. Estimated pK'1 values in NaCl, TC, and TDHC were 6.8, 6.0, and 5.6, respectively; pK'2 was greater than or equal to 9.3 in each system. Acidification of disodium bilirubinate to pH less than 8.5 produced high, metastable [Bt] in 50 mM TC; this was absent in 0.15 M NaCl, and minor in 50 mM TDHC. In all solutions, maximum [Bt] of 60-65 mM was attained at pH greater than or equal to 10.5. This work helps explain the immense variation among reported [Bt] values, indicates that UCB monoanion predominates at the pH range of bile, and suggests that bile salt monomers, dimers, and micelles enhance the solubility of UCB in bile.

Bile Acids and Salts↗
PubMed1987

Increase in a specific cytochrome P-450 isoenzyme in the liver of congenitally jaundiced Gunn rats.

Congenitally jaundiced (jj) Gunn rats had a greater hepatic microsomal content of a cytochrome P-450 isoenzyme, P-450c, than did the non-jaundiced (Jj) rats. No differences in content of P-450b, P-450d and pregnenolone-16 alpha-carbonitrile-induced (PCN) P-450 were found between jj and Jj rats. This is the first demonstration of a constitutive increase in a specific cytochrome P-450 isoenzyme in association with a genetic defect.

Animals↗
PubMed1987

Reliability of balloon-mesh cytology in detecting esophageal carcinoma in a population of US veterans.

A catheter, equipped with a terminal balloon covered with nylon mesh, was developed to study the reliability of abrasive cytology for the diagnosis of esophageal carcinoma. Eighty-seven balloon cytology analyses were attempted in 82 subjects. Four patients were unable to swallow the balloon. In the 78 successful attempts, the initial diagnoses were: esophagitis (34) and esophageal carcinoma (13), established by endoscopic examination and histologic sampling; and normal esophagus (31) confirmed histologically in 25. The remaining 6 controls were younger than 40 years old, without any significant history of smoking, drinking and esophageal symptoms. For esophageal carcinoma, the sensitivity of balloon cytology was 91% and the specificity was 94% with four false-positives. Balloon cytology was generally well-tolerated and easily performed. This method is now being tested for screening high-risk patients for esophageal carcinoma.

Biopsy↗
PubMed1987

Enzymatic oxidation of unconjugated bilirubin to assess its interactions with taurocholate.

The rate of peroxidation of unconjugated bilirubin (UCB), catalyzed by horseradish peroxidase (HRP), has been employed by Jacobsen (1969. FEBS Lett. 5: 112-114) to assess the fraction of unbound UCB in the presence of serum albumin. We used this method to examine the interactions of UCB with taurocholate (TC) at pH 8.2, assuming solubilization of UCB by TC is due to pigment binding and/or to partitioning into the micelle, thus rendering UCB unavailable for peroxidation. Inhibition of UCB peroxidation conformed with predictions based on these assumptions and demonstrated significant interaction of UCB with both monomeric and micellar TC. Although significant inhibition of UCB peroxidation was seen with TC monomer, inhibition was even greater with TC micelles. In contrast, pyrogallol, another substrate of HRP, acted very differently in the presence of TC. Inhibition of pyrogallol peroxidation by TC was much less than with UCB and occurred primarily with monomeric TC, with little further inhibition in the micellar range. The results of this study suggest that at taurocholate concentrations above 50 mM, similar to the physiologic bile salt concentrations in human bile, at least 99% of UCB is bound to bile salt, dramatically decreasing the concentration of unbound UCB. Since bile salts also bind Ca2+, they play a dual role in protection against the precipitation of calcium bilirubinate from bile. Therefore, bile salts are a major factor in the prevention of the formation and growth of pigment gallstones.

Bilirubin↗
PubMed1986

Effects of phototherapy on the activity of microsomal mixed-function monooxygenases and the pharmacokinetics of [14C] hexobarbital in jaundiced and heterozygous Gunn rats.

We assessed the effects of in vivo phototherapy of Gunn rats on the activity of hepatic microsomal mixed-function monoxygenases and on the in vivo pharmacokinetics of [14C]HB. In experiment 1 no serial changes were seen in activities of hexobarbital hydroxylase or benzo(a)pyrene hydroxylase in hepatic microsomes isolated after 2, 4 or 7 days from homozygous jaundiced female Gunn rats exposed to continuous phototherapy or in matched Gunn rats maintained under dim light. In experiment 2 homozygous jaundiced (jj) and heterozygous nonjaundiced (Jj) Gunn rats of both sexes each received i.v. [14C]HB on 2 successive days. In random order, each was exposed on the first or second day to phototherapy for 5.5 hr, beginning 0.5 hr before the administration of HB; otherwise, each was kept under dim light. Plasma [14C]HB in arterial blood samples was separated chromatographically from its labeled metabolites, and biexponential plasma disappearance curves for [14C]HB were analyzed by a SAAM-23 computer program. Clearances in female rats were much slower. In both sexes, the total body clearance and volume of distribution of HB were decreased by 20% during phototherapy of the jj but not the Jj rats; terminal plasma half-life was unchanged. In experiment 3 direct in vitro illumination of [14C]HB did not cause photodegradation of this compound, despite the presence of albumin with or without bilirubin.

Animals↗
PubMed1985

Pruritus in hepatic cholestasis. Pathogenesis and therapy.

Pruritus associated with hepatic cholestasis may cause significant morbidity. Current evidence indicates that bile acids may not be the pruritogenic factor. Rather, the factor may be some other cholephilic anion or anions that bind to cholestyramine resin and are photolabile. Effective treatment modalities used currently include cholestyramine resin and phenobarbital. Phototherapy is a promising modality in which the mechanism and most effective wavelengths remain to be elucidated. Analysis of the possible role of non-steady state bile acid compartmentalization in the pathogenesis of pruritus is presented.

Adrenal Cortex Hormones↗
PubMed1984

Validation of infrared spectroscopy for assessment of vinyl polymers of bile-pigment gallstones.

The i.r. spectra of bilirubin isomers that differ in number and position of vinyl groups were examined to verify the assignment of the 988 cm-1 peak of bilirubin (991 cm-1 peak in calcium bilirubinate) to its pendant vinyl groups. There were only small changes in this peak with changes in position of vinyl groups (exo-2- and -18-vinyl versus endo-3- and -17-vinyl), but progressive loss of peaks in this region was observed when vinyl groups were reduced to ethyl groups (dihydrobilirubin and mesobilirubin). Methylvinylmaleimide, a monopyrrole derived from the outer (A and D) rings of bilirubin, has a pendant vinyl group and exhibits a prominent peak at 986 cm-1, but haematinic acid methyl ester derived from the inner (B and C) rings has no vinyl group and shows no peak near 988 cm-1. These observations verify the assignment of the 988 cm-1 peak of bilirubin to its pendant vinyl groups. This supports our previous proposal that a decrease in the peak at 985-995 cm-1 in the i.r. spectra of pigment gallstones, as compared with unconjugated bilirubin or calcium bilirubinate, indicates a consumption of vinyl groups in the process of formation of the polymer in the pigment stones.

Bilirubin↗
PubMed1984

Polymer networks in pigment and cholesterol gallstones assessed by equilibrium swelling and infrared spectroscopy.

Sixty-five gallstones were assessed for content of network polymer by equilibrium swelling and infrared spectroscopy. All types of pigment gallstones exhibited swelling in aqueous buffers, indicating network polymer content, but swelling was greatest in black pigment gallstones. Gallstones composed principally of cholesterol did not swell. Swelling increased with increasing pH values from 6 to 10.5, suggesting that titratable carboxyl side chains of the pigment were intact. Swelling increased in the presence of 10 mM ethylenediaminetetraacetic acid, indicating that calcium ions provided secondary crosslinks in the network. In all types of pigment gallstones, the equilibrium swelling ratio at the maximum (pH = 10.5) was correlated inversely with the proportion of intact vinyl groups in the pigment assessed by infrared spectroscopy. This suggests that pigment vinyl groups are consumed in forming the covalent crosslinks of the polymer network. A weaker direct correlation between equilibrium swelling ratio and the percentage of unmeasured residue in the gallstones suggested that this residue may also have contributed to the swelling. The high content of cross-linked network polymer in black pigment gallstones renders their therapeutic dissolution improbable by physicochemical means.

Biopolymers↗
PubMed1982

The effect of elimination of environmental light on the metabolism of unconjugated bilirubin in the Gunn rat.

In the homozygous jaundiced Gunn rat, bilirubin catabolism is augmented by intense illumination (phototherapy) and by induction of microsomal cytochrome P448. To assess the relative importance of less intense environmental light versus intrinsic mechanisms in the maintenance of bilirubin turnover, Gunn rats were kept for three weeks under either ordinary laboratory lighting (0.3-0.8 mW/cm2, wavelength range 400-600 nm) or in absolute darkness. No differences in plasma concentration, miscible pool, turnover of bilirubin, or in hepatic cytochrome P448 activity were noted between the two groups over this period. A greater than twofold increase in the biliary excretion of unconjugated bilirubin was noted in the animals maintained under light, but this represented only 2% of total bilirubin turnover. These results suggest that intrinsic(enzymatic ?) pathways are of primary importance in the maintenance of bilirubin metabolism in the glucuronyltransferase-deficient state under ordinary levels of environmental light.

Animals↗