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J D Ostrow

Publications and source records attributed to J D Ostrow.

At least 91 records · Page 5Linked to original sources
PubMed1971

Photocatabolism of labeled bilirubin in the congenitally jaundiced (Gunn) rat.

To elucidate the mechanism by which phototherapy reduces serum bilirubin, studies were performed on the catabolism of labeled bilirubin in homozygous jaundiced Gunn rats before, during, and after a period of exposure to 1700 foot candles of daylight fluorescent light. Following equilibration with the body pool of an intravenously administered tracer dose of (3)H- or (14)C-bilirubin, radioactive and diazo reactive compounds were excreted in the bile at a slow, steady rate and plasma specific activity declined semilogarithmically. Subsequent exposure to light caused a marked increase in the biliary excretion of radioactive and diazoreactive compounds. Fecal and urinary radioactivity increased also but remained minor fractions of the total excreted radioactivity. After extinguishing the lights, these variables reverted gradually to control values. Spectral and chromotographic analysis of the excreted pigments and their azopigments demonstrated that the increased biliary radioactivity during phototherapy consisted of two roughly equal fractions: (a) unconjugated bilirubin, excreted at rates comparable to the output of conjugated bilirubin in the bile of normal nonjaundiced rats; and (b) water-soluble bilirubin derivatives, chromatographically identical with those found in Gunn rat bile under control lighting conditions but different from the products of photodecomposition of bilirubin in vitro. In some animals, phototherapy produced little decline in plasma bilirubin despite comparable acceleration of bilirubin catabolism. This was attributed tentatively to increased synthesis of early labeled bilirubin in these animals.

Animals↗
PubMed1970

Isolation and properties of conjugated bilirubin from bile.

1. A simple, rapid solvent partition method is described for isolation of conjugated bilirubin, free of unconjugated bilirubin, bile salts, phospholipids and cholesterol, from rat bile. Yields are 40-58%. The product is a phosphate-buffered solution containing approx. 0.4mg of bilirubin/ml, principally as mono- and di-glucuronide conjugates. The method may be modified for isolation of conjugates from human bile with 15-22% yield, and for preparation of unconjugated bilirubin from rat or human bile with yields of 55-62%. 2. The conjugated pigment has red-brown fluorescence and an absorption maximum at 450nm with in(mM) 59.8cm(-1). Diazotization by the Malloy-Evelyn method gives a direct Van den Bergh reaction (in water) 12% greater than the total reaction (in methanol), with in(total) 28.4x10(3)lmol(-1)cm(-1) at 550nm. After desalting by elution from Sephadex LH-20 in 50% (v/v) ethanol, the product gave water-soluble mustard-yellow crystalline needles. Such desalted conjugates were precipitated by Pb(2+) but not by Ba(2+), Ca(2+) or Zn(2+). 3. At pH7.0 and 37 degrees C the conjugated bilirubin was oxidized at a rate of 1%/h without hydrolysis, whereas 84% was hydrolysed by beta-glucuronidase or aqueous alkali. 4. Mono- and di-glucuronides were separated by elution from Sephadex LH-20 in 95% (v/v) ethanol or by extraction with chloroform at pH3.2-3.4. The monoconjugated bilirubin did not become labelled during incubation with unconjugated [(14)C]bilirubin, and chromatographed as a single spot without dissociating into unconjugated bilirubin and diglucuronide as would be expected of a complex. 5. After intravenous injection of mono- or di-conjugated [(14)C]bilirubin into normal or Gunn rats, 79-91% was excreted in bile and 2-7% in urine over 2h. In these experiments injected diglucuronide was not hydrolysed whereas 30-41% of injected monoglucuronide was converted into diglucuronide by the normal but not by the Gunn rats. The evidence favours the existence of a true bilirubin mono-glucuronide that is not a complex.

Animals↗
PubMedDate not supplied

Bilirubin chemistry, ionization and solubilization by bile salts.

Bilirubin is a linear tetrapyrrole whose conformation is affected by internal hydrogen bonds formed between the carboxyl side chains and dipyrromethenone rings. Structural variations include: constitutional isomerism of the vinyl or carboxyethyl side chains, geometric isomerism of the methene bridges, tautomerism of the lactam groups, conformational rotations about the central methylene bridge and ionization of one or both carboxyl groups. Aggregation of the dianion into dimers and multimers may occur. The pKa' values of the two carboxyl groups are affected greatly by the environment and may differ widely in micellar solutions like bile. Solubility of bilirubin in water is less than 1 nM at pH = 7 and about 0.1 microM at pH = 8. Nonetheless, it dissolves poorly in most lipid solvents, except for asymmetrical chloroalkanes. Hydrogen bond-breaking solvents, especially dimethyl sulfoxide, are most effective in solubilizing bilirubin. In bile salt solutions, solubility of bilirubin is well above the concentrations of unconjugated bilirubin found in normal human gallbladder bile, and is impaired by lecithin but unaffected by cholesterol. At physiological pH in bile salt solutions, bilirubin is predominantly in its monoanion form that binds readily to the micelles. In such solutions, addition of physiological concentrations of calcium precipitates calcium bilirubinate, leaving residual bilirubin concentrations of up to 15 microM in 50 mM taurocholate or close to the maximum bilirubin concentrations in normal bile. Studies in which disodium bilirubinate is dissolved in bile salt solutions and pH is adjusted to the physiological range reveal that metastable supersaturation with bilirubin may occur and that a mesophase may also form in the presence of lecithin, akin to that seen with cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)

Bile Acids and Salts↗
PubMedDate not supplied

The etiology of pigment gallstones.

Pigment gallstones are of two major types, black and earthy brown, each consisting of calcium salts of bilirubin and other anions, along with an unmeasured residue that is largely mucin glycoproteins. Studies in model systems indicate that the small proportion of unconjugated bilirubin in bile is solubilized by bile salts and that the ionized bilirubin is more soluble than the protonated diacid. Solubility is decreased by added lecithin but is unaffected by cholesterol. At the pH of bile, unconjugated bilirubin exists mainly as a monoanion with sufficient solubility in mixed micelles not to precipitate, were it not for the presence of calcium, which forms highly insoluble salts with unconjugated bilirubin anions. Supersaturation of bile with calcium bilirubinates is inhibited by bile salts, which bind calcium, reducing the activity of free calcium ions. When supersaturation occurs, usually due to increased concentrations of bilirubinate anion, nucleation may be initiated by binding of calcium bilirubinate to mucin glycoproteins in bile. In earthy brown stones, which form mainly in the bile ducts, the pigment is mostly calcium bilirubinate, combined with calcium palmitate. These components form due to hydrolysis, by enzymes in infecting bacteria, of conjugated bilirubin and lecithin, respectively. In black stones, which form mainly in the gallbladder, the pigment is mostly a highly cross-linked network polymer of bilirubin, which is insoluble in all solvents. Concomitant polymerization and oxidation of calcium bilirubinate probably occur in the solid state, after precipitation of the pigment due to hydrolysis of conjugated bilirubin by endogenous beta-glucuronidase from the biliary tract and/or liver.(ABSTRACT TRUNCATED AT 250 WORDS)

Bile↗
PubMedDate not supplied

Effects of phenobarbital on bilirubin metabolism and its response to phototherapy in the jaundiced Gunn rat.

Jaundiced Gunn rats, treated with phenobarbital (60 mg per kg i.p. for 7 to 10 days) showed 25 and 36% decreases in mean plasma bilirubin levels in two experiments (p less than 0.01). Kinetic studies with tracer 14C-bilirubin revealed that there was no change in bilirubin turnover or total pool size due to phenobarbital, but a 49% increase in the hepatic pool and a 27% decrease in the cutaneous pool of bilirubin. The increase in the hepatic pool accounted for over 90% of the bilirubin lost from the plasma. Such pretreatment with phenobarbital did not alter the decline in plasma bilirubin or total bilirubin pool due to subsequent phototherapy. Phenobarbital followed by phototherapy produced a significantly greater reduction in plasma bilirubin levels than either treatment alone. These studies demonstrate that phenobarbital does decrease plasma bilirubin in Gunn rats primarily by shifting the pigment to the liver, and suggests that combined treatment with phenobarbital and phototherapy might be of value in patients with congenital hyperbilirubinemia due to glucuronyl transferase deficiency.

Animals↗
PubMedDate not supplied

Effect of combined treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin and phototherapy on bilirubin metabolism in the jaundiced Gunn rat.

2,3,7,8-Tetrachlorodibenzo-p-dioxin, a potent inducer of microsomal cytochrome P448-dependent monoxygenases, and phototherapy both accelerate bilirubin metabolism and decrease jaundice in Gunn rats. The effects of combined treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin and light were studied in these rats by applying phototherapy for 65 hr, beginning 5 days after induction with 2,3,7,8-tetrachlorodibenzo-p-dioxin. 2,3,7,8-Tetrachlorodibenzo-p-dioxin pretreatment caused a 75% decline in plasma bilirubin in 5 days, with no change thereafter, whether or not the rats were exposed subsequently to phototherapy. In the uninduced rats, plasma bilirubin levels declined by 55% after 40 hr of phototherapy. As determined by [14C]bilirubin kinetics, both 2,3,7,8-tetrachlorodibenzo-p-dioxin and phototherapy increased fractional bilirubin turnover and decreased the total bilirubin pool. In the 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced rats, the contracted bilirubin pool shifted from skin to liver, but these tissue pools did not change further during phototherapy. By contrast, in uninduced rats, phototherapy decreased the cutaneous bilirubin pool, which is the main target of phototherapy. 2,3,7,8-Tetrachlorodibenzo-p-dioxin was more effective than phototherapy in diminishing plasma bilirubin levels and the total bilirubin pool, but the combined treatment (2,3,7,8-tetrachlorodibenzo-p-dioxin followed by phototherapy) was no more effective than 2,3,7,8-tetrachlorodibenzo-p-dioxin alone.

Animals↗