Search PubMed⌕ Search

Biomedical subjects

D M Mock

Publications and source records attributed to D M Mock.

At least 55 records · Page 3Linked to original sources
PubMed1997

Urinary biotin analogs increase in humans during chronic supplementation: the analogs are biotin metabolites.

In human subjects, the metabolic origins of bisnorbiotin and biotin sulfoxide were determined by measuring the urinary excretion of each after chronic administration of large oral doses of biotin. For 2 wk, 14 adult volunteers consumed 1,200 micrograms of biotin per day, an amount approximately 20 times the daily dietary intake. With the use of a high-performance liquid chromatography/avidin-binding assay in untimed urine samples obtained before the first dose of biotin and after the 14th dose, concentrations of biotin, bisnorbiotin, and biotin sulfoxide were measured. Excretion was expressed as concentration ratios to urinary creatinine. Bisnorbiotin and biotin sulfoxide excretion increased 85-fold (P < 0.0001) and 114-fold (P < 0.0001), respectively. The molar percentages of bisnorbiotin and biotin sulfoxide decreased from 28 to 14% (P = 0.006) and from 9 to 5% (P = 0.017), respectively. These data provide evidence that the bisnorbiotin and biotin sulfoxide found in human urine are biotin metabolites. Furthermore, we infer that chronic consumption of large amounts of biotin does not substantially saturate the human biotin pathways of biotransformation.

Administration, Oral↗
PubMed1997

[14C]cyanate labeling of sheep red cells: covalent binding to hemoglobin continues in vivo for a day.

The sheep is a useful model to study fetal and newborn physiology including perinatal erythropoiesis and red cell kinetics. A practical, economical method for measuring red cell survival (RCS) in sheep would be very valuable. However, 51Cr is unsatisfactory, and suitable alternatives have not been published. In the course of investigating [14C]cyanate as a label for sheep red cells, we observed continued covalent labeling over 24 h in vivo that was great enough to introduce a substantial artifact into two commonly used parameters of RCS: posttransfusion recovery (PTR24) and time to 50% decrease (T50) when referenced to time zero. In a simulation of in vivo conditions, the amount of 14C bound to Hb increased 26 +/- 6% (mean +/- 1 SD, n = 11) over 24 h. To investigate the mechanism of the increasing 14C bound, acid-acetone extraction, molecular sieve chromatography, and density gradient separation were used separately or in combination to quantitate intracellular free 14C and 14C covalently bound to intracellular proteins. Free 14C decreased as protein-bound [14C]cyanate increased. These studies provide evidence that covalent binding of [14C]cyanate to intracellular Hb continues in vivo for the first 24 h and that the source of the increase is intracellular free [14C]cyanate. We conclude that 1) PTR24 cannot be accurately determined by [14C]cyanate unless labeled red cells are incubated before infusion to allow the cyanate reaction to approach completion and 2) RCS by [14C]cyanate should be referenced to blood concentrations at 24 h.

Acetone↗
PubMed1997

Circulating red cell volume and red cell survival can be accurately determined in sheep using the [14C]cyanate label.

The sheep commonly serves as an animal model for investigation of human fetal and newborn erythropoiesis and red blood cell kinetics. Measurement of red cell volume (RCV) and survival (RCS) in sheep would be useful for studying mechanisms of neonatal anemia. Unfortunately 51Cr, the standard method for RCV, is not suitable for RCS in sheep because 51Cr leaves the red cell too rapidly. We developed and validated the permanent label [14C]cyanate as a method for measuring both RCV and RCS in sheep. In 19 sheep, RCV was determined simultaneously using [14C]cyanate and 51Cr. RCV determined by [14C]cyanate agreed almost perfectly with RCV by 51Cr; correlation coefficient = 0.990. The line of regression had a slope of 0.94 and an intercept of 40; these parameters are not significantly different from a line of identity. In nine sheep, RCS was determined using [14C]cyanate. Survival after d 1 accurately fit a model containing two components: 1) an early exponential loss likely related to damage caused by labeling and handling and 2) a linear decrease that reflected normal survival of undamaged red cells. Mean potential life span (MPL) determined from the linear phase was 114 +/- 12 d (mean +/- 1 SD). These results agree with reported MPL values determined either by 59Fe or differential hemolysis. Together, these observations establish [14C]cyanate-labeled red cells as a tool for measuring both RCV and RCS in sheep and enhance the value of the ovine model for investigating neonatal anemia.

Anemia↗
PubMed1997

Biotin catabolism is accelerated in adults receiving long-term therapy with anticonvulsants.

Using serum biotin concentration as the indicator, a previous study reported biotin deficiency resulting from long-term anticonvulsant therapy. However, serum biotin may not be a good indicator of tissue biotin status. Using better indicators of biotin status in anticonvulsant-treated subjects, we found increased urinary excretion of biotin catabolites and 3-hydroxyisovaleric acid, an organic acid produced in greater quantities secondary to reduced activity of a biotin-dependent carboxylase. We conclude that anticonvulsant treatment led to increased biotin catabolism and probably to reduced biotin status.

Adolescent↗
PubMed1996

The metabolite profile of radioisotope-labeled biotin in rats indicates that rat biotin metabolism is similar to that in humans.

The egg white-fed rat is the most commonly used animal model of human biotin deficiency. We sought to determine whether the urinary excretion rates of biotin and biotin metabolites in rats are similar to those reported in humans. D-(Carbonyl-14C)biotin was injected intraperitoneally at physiologic doses in 6- to 10-wk-old rats; HPLC and radiometric flow detection were used to specifically identify and quantify biotin and metabolites in urine. Substantial amounts of bisnorbiotin and biotin sulfoxide, the two principal human metabolites, were detected. The excretion rates of biotin and metabolites were strikingly dependent on the dose of biotin. When the dose of [14C]biotin was 30% of the daily dietary intake (a physiologic dose), 50% of the administered 14C was excreted within 24 h; more than half of the excretion was the unchanged vitamin. After d 1, [14C]bisnorbiotin was the major form excreted. For the cumulative 5-d excretion, [14C]biotin accounted for 46 +/- 9%, [14C]bisnorbiotin accounted for 47 +/- 11 %, and [14C]biotin sulfoxide accounted for 8 +/- 4% of the total of biotin, bisnorbiotin, and biotin sulfoxide recovered radioactivity (mean +/- 1 SD, n = 6). These proportions are similar to those reported in humans: biotin = 52 +/- 12%, bisnorbiotin = 35 +/- 9%, and biotin sulfoxide = 13 +/- 4% of total biotin plus metabolites (mean +/- 1 SD, n = 10). Thus, these studies confirm the earlier identification of bisnorbiotin and biotin sulfoxide as the two principal biotin metabolites and provide evidence that the rat is an appropriate model for human biotin metabolism.

Animals↗
PubMed1995

Biotin accounts for only half of the total avidin-binding substances in human serum.

In studies using an avidin-binding assay to measure the serum or plasma concentration of biotin, biotin is sometimes assumed to be equal to the avidin-binding substances detected. To provide a range of values for serum concentrations of biotin, bisnorbiotin, and biotin sulfoxide, HPLC was used to separate avidin-binding substances in human serum, and the chromatographic fractions were assayed for avidin-binding substances (biotin and biotin metabolites). In sera from 15 normal fasting adults, substantial concentrations of avidin-binding substances other than biotin were detected. Two of the principal substances were identified as bisnorbiotin and biotin sulfoxide based on their chromatographic properties. The serum concentrations of bisnorbiotin and biotin sulfoxide varied widely among the individuals. In three subjects, the concentration of bisnorbiotin exceeded that of biotin. The presence of avidin-binding substances in addition to biotin may have confounded previous measurements of the concentration of biotin in serum, plasma, and blood when avidin-binding assays were used. Because bioassay methods for biotin sometimes use organisms for which one or more of these biotin metabolites are growth factors, measurements of biotin in blood using some bioassays are likely to overestimate the concentrations of biotin.

Adult↗
PubMed1994

Requirements for biotin are not affected by the combination of copper deficiency and fructose feeding.

OBJECTIVE: The objective of the present study was to establish whether copper (Cu)-deficient rats fed a diet containing fructose as their sole carbohydrate source require more biotin than the recommended 2 mg/kg diet when egg-white serves as the dietary protein. METHODS: Eighty weanling male Sprague-Dawley rats were randomly divided into 8 groups according to type of dietary carbohydrate (starch or fructose), level of Cu (0.6 micrograms Cu/g diet or 6.0 micrograms Cu/g diet) and level of biotin (2 mg/kg diet or 10 mg/kg diet). RESULTS: Regardless of the level of dietary biotin, Cu-deficient rats fed a fructose-containing diet exhibited growth retardation, anemia, atrophied pancreata, enlarged hearts and similar death rates. The remaining Cu-deficient rats fed fructose were emaciated and sick regardless of dietary biotin levels. The concentration of biotin in serum and biotin content of liver of rats fed fructose were higher than corresponding values from rats fed starch. CONCLUSION: Cu-deficient rats fed fructose are not deficient in biotin compared to published normal values. Supplementation of 10 mg/biotin/kg diet did not improve morbidity or mortality and therefore was not beneficial.

Animals↗
PubMed1994

Biotin uptake by basolateral membrane vesicles of human placenta: normal characteristics and role of ethanol.

This study assessed the mechanism of uptake of biotin by the fetal-facing (basolateral) membrane of the term human placenta. Using membrane vesicles, we showed that most of the uptake was attributable to transfer of the vitamin into the vesicle and that the uptake was saturable, Na-dependent, carrier-mediated, and electroneutral. The rate of uptake was less than for biotin uptake by the maternal-facing (apical) membrane of the human placenta. Because ethanol inhibits biotin uptake by the apical membrane, the effect of ethanol on uptake by basolateral vesicles was investigated. With 10-hr exposure at a concentration of 2 and 3 mg/ml, but not 1 mg/ml, ethanol modestly inhibited biotin uptake. The mechanism of inhibition by alcohol is not known.

Biotin↗
PubMed1993

Biotin and biotin analogs in human urine: biotin accounts for only half of the total.

In studies using avidin-binding assays to measure the urinary excretion of biotin, biotin is sometimes assumed to be equal to the detectable avidin-binding substances present. High performance liquid chromatography was used to separate avidin-binding substances in human urine, and the chromatographic fractions were assayed for avidin-binding substances (biotin and biotin analogs) by a sensitive, specific assay based on binding of biotin to [125I]avidin. In a study of ten normal adults, substantial amounts of avidin-binding substances other than biotin were detected, two of which were bisnorbiotin and biotin sulfoxide. These biotin analogs were initially identified by their chromatographic properties, and identities were confirmed by chemical conversion. The presence of avidin-binding substances in addition to biotin may have confounded previous measurements of the urinary excretion of biotin using avidin-binding assays. Because bioassay methods for biotin often use organisms for which one or more of these biotin analogs are growth factors, measurements of biotin in urine using some bioassay methods are likely to overestimate the concentrations of biotin.

Avidin↗
PubMed1993

Human placental biotin transport: normal characteristics and effect of ethanol.

Biotin, a vitamin essential for many metabolic reactions, is supplied to the fetus exclusively from the mother. Deficiency of biotin in pregnancy leads to impaired fetal growth and development. Alcohol taken in pregnancy likewise may cause fetal growth abnormalities. Normal biotin transport via the placenta and the effects of ethanol on this transport apparently have not been studied. Our aims were to characterize these phenomena for the normal human-term placenta. Using maternal-facing placental membrane vesicles, biotin uptake was sodium- and temperature-dependent, saturable, and inhibited by structural analogs of biotin (desthiobiotin, biocytin, and biotin methyl ester), as well as by 4 and 10 hr exposure to 3 g/liter ethanol. Using the isolated perfused single cotyledon method to measure placental transport of biotin at a perfusion concentration of 1 nM, the overall rate of biotin transport was found to be only 30% that of antipyrine, a freely diffusible marker. Clearance of biotin was approximately 2 ml/hr.g placenta, which was equal to the clearance of passively transferred L-glucose; biotin clearance was similar in both maternal to fetal and fetal to maternal directions. Overall transfer of biotin from maternal to fetal compartments was not inhibited by 500-fold greater concentrations of the three analogs, did not proceed against a biotin concentration gradient, and was not inhibited by 90-240 min exposure to an initial concentration of 4 g/liter ethanol. Concentration of biotin in the fetal compartment at the end of the study was not higher than on the maternal side (after maternal to fetal infusion), but placental concentration was 2- to 3-fold greater. No significant metabolism of biotin was detected. Exposing human placental cultured trophoblast on day 3 to 24 hr of ethanol (2 g/liter) had no effect on the net uptake of biotin by these cells. These studies provide evidence that maternal-facing placental membranes take up biotin by a mediated, carrier-dependent process that is inhibited by ethanol; however, based on the perfusion studies, we conclude that the overall (maternal-fetal) rate-limiting transfer of biotin by the human placenta is most consistent with a passive process, which is not inhibited by short-term exposure to ethanol.

Biotin↗
PubMed1992

Rayon balls and disposable-diaper material selectively adsorb creatinine.

One method for the collection of urine samples from infants involves absorption of the urine on cotton balls placed in the diaper. Such samples are not timed and excretions are often expressed per mg of urinary creatinine. An assumption in this method is that the creatinine concentration is not changed by the absorption process. The concentration of creatinine in urine samples was measured before and after absorption of the urine by cotton balls, rayon balls, or diaper material over a range of wetness. For urine from both adults and infants, absorption on rayon balls and diaper material caused an important artifactual decrease in the concentration of creatinine. The effect was particularly striking in lightly wetted samples; the mean percent decrease was only 3 +/- 2% for cotton but was 9 +/- 5% for rayon (n = 10) and 13 +/- 4% for diaper material (n = 7). These data provide evidence that rayon balls and diaper material (and to a lesser extent cotton balls) selectively adsorb creatinine from human urine.

Adsorption↗
PubMed1992

Distribution of biotin in human plasma: most of the biotin is not bound to protein.

Estimates of the plasma concentration of biotin differ considerably. Variation in detectability of biotin bound covalently to protein is one potential source of disagreement. In this study we determined the amount of biotin covalently bound to plasma protein. First, greater than 99% of free and reversibly bound biotin was removed by dialysis; then greater than 90% of covalently bound biotin was released by acid hydrolysis. For plasma samples from 11 normal adults, the ratio of covalently bound biotin to free biotin was 0.15 +/- 0.09 (mean +/- SD). Taking into account the additional biotin that is reversibly bound to protein, this study provides evidence that approximately 12% of total biotin in plasma is covalently bound, 7% is reversibly bound, and 81% is free. We conclude that covalently bound biotin cannot account for the reported sixfold increase in biotin detected after acid hydrolysis. We speculate that the reported increase was an artifact caused by substances produced during acid hydrolysis of plasma.

Adult↗
PubMed1992

Biotin in human milk: methods, location, and chemical form.

For infants, no Recommended Dietary Allowance for biotin has been published; the estimated safe and adequate intake seems to be based on measurements of human milk. However, published estimates of the biotin content disagree substantially. We sought to address several of the potential sources of disagreement by defining the conditions for the collection, storage, and subcompartment distribution of biotin in human milk using the [125I]avidin assay. Composition of the collection vessel (glass vs. common plastics) had no effect on biotin content of human milk. The biotin content of milk did not change during storage at room temperature for at least 1 wk, at 5 degrees C for at least 1 mo, or at -20 degrees C or -70 degrees C for at least 1.5 y. Biotin in the cell pellet and fat fraction accounted for less than 5% of that in the skim fraction. Of the biotin in the skim fraction, none (less than 3%) was reversibly bound to macromolecules, and less than 5% was covalently bound to macromolecules. We conclude that assay of free biotin will detect almost all of the biotin present in a sample of mature human milk.

Adult↗
PubMed1992

Secretory patterns of biotin in human milk.

The current recommendation for safe and adequate daily dietary intake of biotin for infants is based on measurements of biotin concentration in human milk and calculations of biotin intake that tacitly assume that biotin content of human milk is reasonably uniform for a given subject. This assumption of uniformity was tested by examining the effects of several factors on the biotin concentration. The degree of breast emptying had little effect on biotin concentration. However, in three of five individuals studied, the biotin concentration varied significantly over 24 h. In two of five subjects, there was a consistent difference between breasts of approximately 16%. In the first 18 d postpartum, the milk concentration of biotin increased steadily in four of the eight individuals studied, remained low in two and increased erratically in two. Rather than reaching a stable plateau in mature milk, biotin concentration varied substantially in most of the subjects. These observations provide evidence that an adequate scheme for estimating total biotin intake of the breastfed infant will require sampling from both breasts frequently over the 24-h cycle and frequently as a function of time postpartum.

Analysis of Variance↗
PubMed1992

Biotin deficiency in rats: disturbances of leucine metabolism are detectable early.

3-Methylcrotonyl-CoA originates from catabolism of leucine and is normally metabolized to acetyl-CoA. However, in biotin deficiency, reduced hepatic activity of the biotin-dependent enzyme methylcrotonyl-CoA carboxylase causes the enzyme's substrate 3-methylcrotonyl-CoA to be shunted via an alternate pathway to 3-hydroxyisovaleric acid (3-HIA), which is excreted at increased rates in the urine. In a previous study, unequivocal separation in 3-HIA excretion rates between biotin-deficient and control animals was not apparent until d 35 of feeding a diet that induced biotin deficiency. The present study tested the hypothesis that abnormal 3-HIA excretion could be detected earlier in the course of biotin deficiency if 3-HIA were more accurately measured using a method that incorporated an improved extraction regimen, deuterated 3-HIA as internal standard, and unlabeled 3-HIA as external standard. Biotin deficiency was induced in rats by feeding a diet containing avidin; control rats received the same diet and biotin injections. With the more accurate method, unequivocal detection of deficiency was possible in all deficient rats by d 16. This study provides evidence that, in rats, reduction of analytical error allows earlier detection of biotin deficiency and that disturbances of leucine metabolism occur earlier than previously appreciated.

Animals↗
PubMed1991

Skin manifestations of biotin deficiency.

This article reviews current knowledge concerning the dermatologic manifestations of biotin deficiency. Biotin is a water-soluble vitamin that acts as an essential cofactor for four carboxylases, each of which catalyzes an essential step in intermediary metabolism. For example, acetyl-CoA carboxylase catalyzes the rate-limiting step in fatty acid elongation. In infants, children, and adults, deficiency of biotin causes alopecia and a characteristic scaly, erythematous dermatitis distributed around body orifices. The rash closely resembles that of zinc deficiency. Candida albicans often can be cultured from the skin lesions. Biotinidase deficiency, an inborn error, causes biotin deficiency, probably as a consequence of unpaired intestinal absorption, cellular salvage, and renal reclamation of biotin; biotinidase deficiency causes dermatologic manifestations similar to biotin deficiency. There is evidence that impaired fatty acid metabolism secondary to reduced activities of the biotin-dependent carboxylases (especially acetyl-CoA carboxylase) plays an etiologic role in the dermatologic manifestations of biotin deficiency. Candida infections secondary to impaired immune function might also contribute to the dermatitis of biotin deficiency.

Adult↗