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R L Horst

Publications and source records attributed to R L Horst.

At least 163 records · Page 9Linked to original sources
PubMed1983

Intestinal synthesis of 24-keto-1,25-dihydroxyvitamin D3. A metabolite formed in vivo with high affinity for the vitamin D cytosolic receptor.

24-Keto-1,25-dihydroxyvitamin D3 has been identified as an intestinal metabolite of 1,25-dihydroxyvitamin D3 by ultraviolet absorbance, mass spectroscopy, and chemical reactivity. The metabolite was produced from 1,25-dihydroxyvitamin D3 and 1,24R,25-trihydroxyvitamin D3 in rat intestinal mucosa homogenates. 24-Keto-1,25-dihydroxyvitamin D3 is present in vivo in the plasma and small intestinal mucosa of rats fed a stock diet, receiving no exogenous 1,25-dihydroxyvitamin D3, and in the plasma and small intestinal mucosa of rats dosed chronically with 1,25-dihydroxyvitamin D3. 24-Keto-1,25-dihydroxyvitamin D3 has affinity equivalent to 1,24R,25-trihydroxyvitamin D3 for the 3.7 S cytosolic receptor specific for 1,25-dihydroxyvitamin D3 in the intestine and thymus. In cytosolic preparations contaminated with the 5 S vitamin D-binding protein, both metabolites are about 7-fold less potent than 1,25-dihydroxyvitamin D3. In contrast, in cytosolic preparations largely free of the 5 S binding protein, both metabolites are equipotent with the parent compound. No evidence was obtained supporting a substantial presence of 23-keto-1,25-dihydroxyvitamin D3 in vivo; nor was the latter compound generated in detectable amounts from 1,25-dihydroxyvitamin D3 by intestinal homogenates. Thus, C-24 oxidation is a significant pathway of intestinal 1,25-dihydroxyvitamin D3 metabolism that produces metabolites with high affinity for the cytosolic receptor which mediates vitamin D action.

Animals↗
PubMed1983

19-nor-10-ketovitamin D derivatives: unique metabolites of vitamin D3, vitamin D2, and 25-hydroxyvitamin D3.

Three metabolites were isolated after incubation of vitamin D3, vitamin D2, or 25-hydroxyvitamin D3 with bovine rumen microbes. They are identified as 5(E)-19-nor-10-ketovitamin D3, 5(E)-19-nor-10-ketovitamin D2, and 5(E)-19-nor-10-keto-25-hydroxyvitamin D3, respectively. The identifications were based on ultraviolet absorbance, mass spectroscopy, and chemical reactivity. All 5(E)-19-nor-10-ketovitamin D derivatives examined had an absorbance maximum at 312 nm and a characteristic fragment in their mass spectra corresponding to loss of 43 amu from their molecular ions. The vitamin D3 metabolite was identical in all essential spectral and chromatographic aspects with authentic synthetic 5(E)-19-nor-10-ketovitamin D3. These metabolites represent a unique pathway of vitamin D metabolism and the first characterized products of microbial vitamin D metabolism. The conversion of vitamin D and its metabolites to their 19-nor-10-keto forms likely represents a detoxification mechanism.

Animals↗
PubMed1983

(23S)-1,23,25-Trihydroxycholecalciferol, an intestinal metabolite of 1,25-dihydroxycholecalciferol.

(23S)-23,25-Dihydroxycholecalciferol was converted into a polar metabolite in a calciferol-deficient chick kidney homogenate. The metabolite was identified as (23S)-1,23,25-trihydroxycholecalciferol by absorbance spectroscopy and mass spectrometry, and by formation of derivatives. (23S)-1,23,25-Trihydroxycholecalciferol was also observed as a 1,25-dihydroxycholecalciferol metabolite in intestinal cells isolated from 1,25-dihydroxycholecalciferol-treated rat. The trihydroxy metabolite was 50-fold less potent than 1,25-dihydroxycholecalciferol in the chick intestinal 1,25-dihydroxycholecalciferol receptor assay.

Animals↗
PubMed1983

23-Keto-25-hydroxyvitamin D3: a vitamin D3 metabolite with high affinity for the 1,25-dihydroxyvitamin D specific cytosol receptor.

A new metabolite of 23,25-dihydroxyvitamin D3 has been generated with kidney homogenates prepared from vitamin D treated chicks. The metabolite was purified with three high-performance liquid chromatographic steps and was identified as 23-keto-25-hydroxyvitamin D3 by ultraviolet absorption spectroscopy, mass spectrometry, and chemical reactivity. The R stereoisomer of 23,25-dihydroxyvitamin D3 was 10-fold more effective as an in vitro precursor to 23-keto-25-hydroxyvitamin D3 than was the naturally occurring S stereoisomer. Approximately 500 ng of 23-keto-25-hydroxyvitamin D3 was necessary to produce the same degree of intestinal-calcium transport as 25 ng of vitamin D3--a difference of about 20-fold. 23-Keto-25-hydroxyvitamin D3 was not active at stimulating bone calcium resorption at the doses and times tested. This new vitamin D3 metabolite, however, had greater affinity than 25-hydroxyvitamin D3 to both the rat plasma vitamin D binding protein and the 1,25-dihydroxyvitamin D specific cytosol receptor. Heretofore, only 1 alpha-hydroxylated metabolites of 25-hydroxyvitamin D3 or analogues possessing a pseudo 1 alpha-hydroxy group were known to bind to the 1,25-dihydroxyvitamin D receptor with higher affinity than 25-hydroxyvitamin D3. Ketone formation at the 23 position, therefore, is the first side-chain modification of 25-hydroxyvitamin D3 that results in enhanced binding to the 1,25-dihydroxyvitamin D receptor binding protein.

Animals↗
PubMed1983

Metabolism of orally administered [3H]ergocalciferol and [3H]cholecalciferol by dairy calves.

Concentrations of ergocalciferol, cholecalciferol, and their metabolites in plasma were determined after a single oral dose of [3H]ergocalciferol or [3H]cholecalciferol was given to 95- to 105-kg Jersey bull calves. One group (three calves) was given 365 muCi of [3H]ergocalciferol (1.2 Ci/mmol) per calf, and the other group (three calves) was given 365 muCi of [3H]cholecalciferol (1.2 Ci/mmol) per calf. Fourteen blood samples were taken from each calf during the 3 weeks after administration. Total plasma radioactivity was highest at 80 hours in both groups (8400 dpm/ml and 4600 dpm/ml in the [3H]cholecalciferol- and [3H]ergocalciferol-treated calves, respectively). For determination of the time-dependent appearance and disappearance of plasma vitamin D and vitamin D metabolites, the plasma 3H-labeled steroids were extracted and separated by high-performance liquid chromatography. In both groups, [3H]vitamin D peaked at 24-48 hours and was the predominant radioactive form in plasma 10-15 hours after dosing. After 15 hours, 25-[3H]hydroxyvitamin D became the predominant labeled metabolite, reaching its maximal concentration between 48 and 96 hours. Concentrations of 25-[3H]hydroxycholecalciferol were about twice those of 25-[3H]hydroxyergocalciferol. The appearance/disappearance profile of 25,26-[3H]dihydroxycholecalciferol and 1,25[3H]hydroxycholecalciferol resembled that of 25-[3H]hydroxycholecalciferol.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed1983

A detailed evaluation of oral phosphate therapy in selected patients with primary hyperparathyroidism.

Recent studies have emphasized the pathophysiological importance of circulating 1,25-dihydroxyvitamin D ((1,25-(OH)2D] in the pathogenesis of hypercalciuria and renal stone formation in primary hyperparathyroidism. Reasoning that phosphate administration might be capable of reducing the plasma concentration of 1,25-(OH)2D in patients with a prominent 1,25-(OH)2D-mediated absorptive component to their disease, 10 carefully selected patients were treated with oral phosphate (1500 mg elemental phosphorus daily) for 1 yr. Phosphate treatment significantly reduced circulating 1,25-(OH)2D levels (84 to 56 pg/ml), the calciuric response to an oral calcium tolerance test (0.30 to 0.21 delta mg calcium/dl GF), and calcium excretion on an unrestricted calcium diet (438-269 mg/day), in essence reversing the absorptive pattern of abnormalities observed before treatment. This response, however, was accompanied by an increase in biochemical hyperparathyroidism, as assessed by circulating immunoreactive PTH and nephrogenous cAMP excretion. In patients with biochemical evidence of an increase in bone resorption before therapy, histomorphometric, radiographic, and biochemical data revealed a trend toward a reduction in bone turnover during phosphorus therapy, with an apparent maintenance of coupled bone resorption and bone formation. This trend, however, was of marginal statistical significance in the patient group as a whole. It is concluded 1) that phosphate therapy represents a viable medical alternative in selected patients with primary hyperparathyroidism, 2) that the net response in treated patients is multifaceted and complex, and 3) that the efficacy of phosphate therapy will ultimately depend upon its long term effects on skeletal homeostasis.

Bone and Bones↗
PubMed1983

Effects of 1 alpha-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3, 1,24,25-trihydroxyvitamin D3, and 1,25,26-trihydroxyvitamin D3 on mineral metabolism and 1,25-dihydroxyvitamin D concentrations in dairy cows.

Six mature, nonlactating, nonpregnant Jersey cows were treated with five 1-alpha-hydroxylated metabolites of vitamin D3 to compare relative potencies of the metabolites. Blood plasma 1,25-dihydroxyvitamin D peaked at about 1000 pg/ml 12 h after intramuscular injection of 500 micrograms 1,25-dihydroxyvitamin D3 but returned to preinjection concentration within 3 to 4 days. Administration of 500 micrograms 1-alpha-hydroxyvitamin D3 intramuscularly led to a gradual increase in 1,25-dihydroxyvitamin D concentration to 500 pg/ml 48 h after treatment. When 500 micrograms of 1,25-dihydroxyvitamin D3 or 1 alpha-hydroxyvitamin D3 were given orally, plasma 1,25-dihydroxyvitamin D peaked at about 200 and 100 pg/ml, respectively. The decline from peak concentrations occurred more slowly after oral treatment than after intramuscular treatment. Plasma calcium was increased by 20 to 30% for a week or more, depending on 1,25-dihydroxyvitamin D concentrations in plasma. All treatments resulted in calciuria. Treatment with 1 or 3 mg doses of 1,24(R),25-trihydroxyvitamin D3 or both carbon-25 stereoisomers of 1,25,26-trihydroxyvitamin D3 induced smaller and less protracted changes in plasma minerals.

Administration, Oral↗
PubMed1983

Plasma concentrations of 1,25-dihydroxyvitamin D, 1,24R,25-trihydroxyvitamin D3, and 2,25,26-trihydroxyvitamin D3 after their administration to dairy cows.

Intramuscular administration of 1,25 (R or S),26-trihydroxyvitamin D3 to dairy cows resulted in significant elevation of 1,25-dihydroxyvitamin D3 concentration in blood plasma. The increase of 1,25-dihydroxyvitamin D3 in plasma paralleled the increase of 1,25,26-trihydroxyvitamin D3. We also found that intramuscular 1,25-dihydroxyvitamin D3 administration to dairy cows significantly elevated plasma 1,24,25-trihydroxyvitamin D3. These data suggest possible impairment of 1,25-dihydroxyvitamin D3 metabolism during the presence of pharmacologic concentrations of 1,25,26-dihydroxyvitamin D3 in plasma. In addition, the 24-hydroxylation of 1,25-dihydroxyvitamin D3 to 1,24,25-trihydroxyvitamin D3 appears to be an important route for further metabolism of 1,25-dihydroxyvitamin D3 in dairy cows.

Animals↗
PubMed1982

23S,25-dihydroxyvitamin D3 as a circulating metabolite of vitamin D3. Its role in 25-hydroxyvitamin D3-26,23-lactone biosynthesis.

23,25-Dihydroxyvitamin D3 was isolated from vitamin D-toxic pig plasma by sequential chromatography through two gravity columns and three high performance liquid chromatography systems. Two of the high performance liquid chromatography systems separated the R and S diastereomers of 23,25-dihydroxyvitamin D3 and demonstrated that the metabolite has the 23S configuration. Ultraviolet absorbance and mass spectroscopy of the pure metabolite and mass spectroscopy of its trisilylated derivative confirmed the structural assignment. 23S,25-Dihydroxyvitamin D3, 23R,25-dihydroxyvitamin D3, 25S,26-dihydroxyvitamin D3, and 25R,26-dihydroxyvitamin D3 were assessed for their ability to produce 25-hydroxyvitamin D3-26,23-lactone in vitamin D2-toxic rats. On 23S,25-dihydroxyvitamin D3, of the naturally occurring compounds, was able to increase the plasma lactone concentration. This metabolite was a more efficient precursor than 25-hydroxyvitamin D3, suggesting that 23S-hydroxylation is a rate-limiting step in 25-hydroxyvitamin D3-26,23-lactone formation. 23S,25-Dihydroxyvitamin D3 was not detected in 25-hydroxyvitamin D3-dosed rats, indicating that the former is rapidly metabolized. Nephrectomized rats had a diminished but significant ability to synthesize 25-hydroxyvitamin D3-26,23-lactone from 25-hydroxyvitamin D3. Nephrectomy did not affect synthesis of 25-hydroxyvitamin D3-26,23-lactone from 23S,25-dihydroxyvitamin D3. These results demonstrate that vitamin D3 23S-hydroxylase(s) are also located extrarenally and that extrarenal tissues are quantitatively important to 25-hydroxyvitamin D3-26,23-lactone synthesis.

Animals↗
PubMed1982

23, 25, 26-trihydroxycholecalciferol. A 25-hydroxycholecalciferol-26,23-lactone precursor.

(23S)-23,25-Dihydroxycholecalciferol was converted into at least five metabolites in kidney homogenates prepared from 1,25-dihydroxycholecalciferol-treated chickens. One of these has been positively identified as 23,25,26-trihydroxycholecalciferol by u.v.-absorbance analysis, mass spectrometry and chemical formation of derivatives. 23,25,26-Trihydroxycholecaciferol produces 25-hydroxycholecalciferol-26,23-lactone when incubated in chick kidney homogenates.

Animals↗
PubMed1982

Discrimination in the metabolism of orally dosed ergocalciferol and cholecalciferol by the pig, rat and chick.

Vitamin D-deficient pigs, rats and chicks were repleted with four daily oral doses of crystalline ergocalciferol (vitamin D2) and cholecalciferol (vitamin D3) containing equal concentrations of each. At 24 h after the last dose, the plasma of each species was analysed for vitamin D and 25-hydroxyvitamin D by standard methods. The mean (+/- S.D.) ratios of plasma cholecalciferol to ergocalciferol concentration were 1.5 +/- 0.1 (pig). 1.7 +/- 0.1 (rat) and 6.3 +/- 1.2 (chick). The mean ratios of plasma 25-hydroxycholecalciferol to 25-hydroxyergocalciferol concentration were 4.0 +/- 0.1 (pig), 0.4 +/- 0.02 (rat) and 10.7 +/- 3.4 (chick). The mean plasma cholecalciferol/ergocalciferol ratios for the 24,25-dihydroxy-, 25,26-dihydroxy- and 1,25-dihydroxy-derivatives in the pig were 2.6 +/- 0.6, 5.8 +/- 1.3 and 5.8 +/- 0.8 respectively. This is the first evidence that mammals other than the New World monkey, like birds, discriminate between ergocalciferol and cholecalciferol. These data, therefore, suggest that species discrimination between the different forms of vitamin D is probably a general phenomenon in mammals. Moreover, this is the first indication of a species (rat) that discriminates against a cholecalciferol metabolite in favour of an ergocalciferol metabolite. Species discrimination against particular forms of vitamin D may be important to the choice of experimental animal models for studying the regulation of vitamin D metabolism and may also be an important consideration in dietary vitamin supplementation.

25-Hydroxyvitamin D 2↗
PubMed1982

Comparison of plasma concentrations of vitamin D and its metabolites in young and aged domestic animals.

1. Vitamin D and its metabolites were measured in the plasma of five species of rural domestic animals. 2. Concentration of 1,25-dihydroxyvitamin D was higher (P less than 0.01) in young animals (range 24-118 pg/ml, means +/- SD = 72.0 +/- 30.0) than in adult animals (range 14-67 pg/ml, means +/- SD = 40.2 +/- 22.6). 3. 25-Hydroxyvitamin D3-26,23 lactone was present only in the chick and the pig. 4. Unsheared sheep appeared to be inefficient utilizers of the photochemical conversion of 7-dehydrocholesterol to vitamin D3. 5. Conversion of 25-hydroxyvitamin D to 24,25-dihydroxyvitamin D was most efficient in species with high plasma phosphorus concentrations (pig, sheep).

24,25-Dihydroxyvitamin D 3↗