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Biomedical subjects

R P Holmes

Publications and source records attributed to R P Holmes.

At least 19 recordsLinked to original sources
PubMed1999

Guaifenesin- and ephedrine-induced stones.

PURPOSE: We report a new type of drug-induced stone that is caused by overconsumption of preparations containing guaifenesin and ephedrine. MATERIALS AND METHODS: Clinical and stone analysis data from the Molecular Structure Laboratory at the Veterans Affairs Medical Center in Milwaukee, Wisconsin, were reviewed. Stone analysis was performed by Fourier transform infrared spectroscopy, high-resolution X-ray crystallographic powder diffraction, or both. The urine and stone material from one of the subjects were analyzed with high-performance liquid chromatography. RESULTS: Stone analysis from seven patients demonstrated metabolites of guaifenesin. High-performance liquid chromatography revealed that the stone and urine from one subject had a high content of guaifenesin metabolites and a small amount of ephedrine. Demographic data were available on five patients. Three had a history of alcohol or drug dependency. All were consuming over-the-counter preparations containing ephedrine and guaifenesin. Four admitted to taking excessive quantities of these agents, mainly as a stimulant. Hypocitraturia was identified in two individuals subjected to urinary metabolic testing. These stones are radiolucent on standard X-ray imaging but can be demonstrated on unenhanced CT. Shockwave lithotripsy was performed in two patients, and the calculi fragmented easily. CONCLUSIONS: Individuals consuming large quantities of preparations containing ephedrine and guaifenesin may be at risk to develop stones derived mainly from metabolites of guaifenesin and small quantities of ephedrine. These patients may be prone to drug or alcohol dependency.

Adult↗
PubMed1999

The gene encoding hydroxypyruvate reductase (GRHPR) is mutated in patients with primary hyperoxaluria type II.

Primary hyperoxaluria type II (PH2) is a rare monogenic disorder that is characterized by a lack of the enzyme that catalyzes the reduction of hydroxypyruvate to D-glycerate, the reduction of glyoxylate to glycolate and the oxidation of D-glycerate to hydroxypyruvate. The disease is characterized by an elevated urinary excretion of oxalate and L-glycerate. The increased oxalate excretion can cause nephrolithiasis and nephrocalci-nosis and can, in some cases, result in renal failure and systemic oxalate deposition. We identified a glyoxylate reductase/hydroxypyruvate reductase (GRHPR) cDNA clone from a human liver expressed sequence tag (EST) library. Nucleotide sequence analysis identified a 1198 nucleotide clone that encoded a 984 nucleotide open reading frame. The open reading frame encodes a predicted 328 amino acid protein with a mass of 35 563 Da. Transient transfection of the cDNA clone into COS cells verified that it encoded an enzyme with hydroxy-pyruvate reductase, glyoxylate reductase and D-glycerate dehydrogenase enzymatic activities. Database analysis of human ESTs reveals widespread tissue expression, indicating that the enzyme may have a previously unrecognized role in metabolism. The genomic structure of the human GRHPR gene was determined and contains nine exons and eight introns and spans approximately 9 kb pericentromeric on chromosome 9. Four PH2 patients representing two pairs of siblings from two unrelated families were analyzed for mutations in GRHPR by single strand conformation polymorphism analysis. All four patients were homozygous for a single nucleotide deletion at codon 35 in exon 2, resulting in a premature stop codon at codon 45. The cDNA that we have identified represents the first characterization of an animal GRHPR sequence. The data we present will facilitate future genetic testing to confirm the clinical diagnosis of PH2. These data will also facilitate heterozygote testing and prenatal testing in families affected with PH2 to aid in genetic counseling.

Alcohol Oxidoreductases↗
PubMed1999

Maternal serum insulin-like growth factor binding protein-2 and -3 and fetal growth.

This was a prospective observational study of maternal insulin-like growth factor binding protein-2 and -3 and fetal growth in 141 pregnant women after 24 weeks gestation who were scanned and venesected fortnightly. Cases (birthweight <5th centile) were sub-divided into those with growth restriction due to placental dysfunction (n = 25) and normal small (n = 27) and there were 89 normally grown controls. Maternal binding protein-3 was measured by radioimmunoassay and the overall pattern of the binding proteins and their proteolytic modifications were assessed by Western ligand blotting and immunoblotting followed by densitometric analysis. In controls, there was no correlation between binding protein-3 and birthweight, and binding protein-3 was elevated in the normal small but not the placental dysfunction group. Complete proteolysis of the 40 kDa doublet of binding protein-3 was observed in all pregnancies. Maternal serum binding protein-2 concentrations were unchanged in normal pregnancy compared to non-pregnant controls but elevated in the growth-restricted group and in all pregnancies binding protein-2 was predominantly present as a 14 kDa proteolysed fragment. These results suggest that compensatory changes in binding protein-2 and -3 or their proteolysis do not increase bioavailability and so do not confound the low maternal insulin-like growth factor-I in growth restricted pregnancies.

Case-Control Studies↗
PubMed1999

Direct quantification of the enteric bacterium Oxalobacter formigenes in human fecal samples by quantitative competitive-template PCR.

Homeostasis of oxalic acid appears to be regulated, in part, by the gut-associated bacterium Oxalobacter formigenes. The loss of this bacterium from the gut flora is associated with an increased susceptibility to hyperoxaluria, a condition which can lead to the formation of calcium oxalate crystalluria and kidney stones. In order to identify and quantify the presence of O. formigenes in clinical specimens, a quantitative-PCR-based assay system utilizing a competitive DNA template as an internal standard was developed. This quantitative competitive-template PCR test allows for the rapid, highly specific, and reproducible quantification of O. formigenes in fecal samples and provides a prototype for development of DNA-based quantitative assays for enteric bacteria.

Bacteria↗
PubMed1999

Glycolate metabolism by Hep G2 cells.

The pathways of oxalate synthesis in humans are not well defined despite their clinical significance in primary hyperoxaluria and idiopathic calcium oxalate nephrolithiasis. Furthermore, the functional roles, if any, of this synthesis have not been elucidated. This study examines pathways of oxalate synthesis from glycolate in Hep G2 cells, a human hepatoma cell line. Incubation of these cells with glycolate has revealed that a pathway may function to synthesize oxalate from glycolate that does not depend on the oxidation of glycolate to glyoxylate by glycolate oxidase. Labeling cells with 14C-glycolate and chromatographic analyses indicated that detectable amounts of 14C-glyoxylate were not formed. A radioactive peak that coeluted with oxalate on ion exclusion chromatography was the only peak yet identified. A detailed examination of glycolate metabolism in these cells should help clarify the terminal steps associated with oxalate synthesis and aid in our understanding of two-carbon metabolism.

Carcinoma, Hepatocellular↗
PubMed1998

Genetic and dietary influences on urinary oxalate excretion.

Several genes contribute to the development of calcium oxalate nephrolithiasis as it is a polygenic disease. To explore the influence of genetic factors on oxalate excretion we have examined the distribution of oxalate excretions in 101 normal individuals who consumed self-selected diets. The distribution was apparently trimodal, consistent with the existence of three classes of oxalate excretors reflecting two allelic genes determining high and low oxalate excretion occurring with frequencies of 0.32 and 0.68 respectively. The pattern of inheritance in eight families was compatible with the expression of a pair of codominant alleles. A comparison of the distribution of excretory classes among the 101 normal individuals with that of 101 calcium oxalate stone formers suggests that high oxalate excretion may be associated with a 4-fold increased risk of stone disease and intermediate excretion with a 1.6-fold increase. Control of dietary factors influencing oxalate excretion apparently improved the discrimination between excretory classes in 17 individuals but the intra-individual variability in oxalate excretion was not reduced in four of nine individuals in whom this parameter was evaluated. More stringent dietary control than that applied in this study may be required before more extensive genotyping of individuals is attempted.

Adult↗
PubMed1998

A prospective study of maternal serum insulin-like growth factor-I in pregnancies with appropriately grown or growth restricted fetuses.

OBJECTIVE: To determine whether there is a relationship between maternal serum insulin-like growth factor-I and fetal growth, consistent with the hypothesis that insulin-like growth factor-I influences maternal constraint upon fetal growth by controlling placental transfer. DESIGN: A prospective, observational study. SETTING: Fetal medicine unit and antenatal clinic of a large teaching hospital. POPULATION: One hundred and forty-one pregnant women identified as having small or normally grown fetuses. METHODS: Fetuses were scanned every two weeks with maternal venesection at each visit. Cases (birthweight < 5th centile) were assigned to two groups: fetal growth restriction due to placental dysfunction (umbilical artery Doppler, growth velocity pulsatility index > +2 SD; n = 25) and normal small-for-gestational-age (normal Doppler, growth velocity and amniotic fluid; n = 27). Eighty-nine controls had birthweights between the 5th and the 95th centiles, normal Doppler, growth velocity and amniotic fluid. Insulin-like growth factor-I was measured by radioimmunoassay, and its relationship to gestational age and birthweight was assessed by regression analysis. Comparisons between case groups were made by Student's t test or analysis of covariance to allow for the effect of birthweight. OUTCOME MEASURE: The last insulin-like growth factor-I level before delivery within the different subgroups. RESULTS: In controls, maternal insulin-like growth factor-I increased with gestational age (r = 0.40; P = 0.0001) but did not correlate with birthweight. Insulin-like growth factor-I was low in the mothers of growth restricted fetuses (-1.56 SD; P = 0.0001), but not in those with small-for-gestational age fetuses. CONCLUSIONS: The control and small-for-gestational-age data suggest that maternal insulin-like growth factor-I is not associated with endocrine control of normal placental function. Low insulin-like growth factor-I relates to poor placental transfer, as indicated by Doppler, rather than to low birthweight. Whether this is a regulatory mechanism, a cause or a consequence of placental dysfunction needs further study.

Biomarkers↗
PubMed1998

Glyoxylate synthesis, and its modulation and influence on oxalate synthesis.

PURPOSE: We define the major pathways of hepatic oxalate synthesis in humans, examine the association with other metabolic pathways and identify ways that oxalate synthesis may be modified. In addition, we suggest what is required for further progress in this area. MATERIALS AND METHODS: We consolidated relevant data primarily from recently published literature, considered new pharmacological approaches to decrease oxalate synthesis, and formulated an overview of the regulation and modification of oxalate synthesis pathways. RESULTS: Experiments with animals, including humans, animal cells and in vitro preparations of cellular components, support the existence of a major metabolic pathway linking the amino acids serine, glycine and alanine. Oxalate synthesis is a minor, secondary reaction of a cascade of reactions termed the glyoxylate pathway, which has a prominent role in gluconeogenesis and ureagenesis. The enzymatic steps and effectors which regulate glyoxylate and oxalate synthesis are not well characterized. Pharmacological approaches can reduce oxalate synthesis by diminishing the glyoxylate pool and possibly modifying enzymatic reactions leading to glyoxylate synthesis. CONCLUSIONS: The individual steps associated with glyoxylate and oxalate synthesis can be identified. The glyoxylate pathway has a significant functional role in intermediary liver metabolism but the way it is regulated is uncertain. Oxalate synthesis can be modified by drugs, indicating that primary and idiopathic hyperoxaluria may respond to pharmacological intervention.

Animals↗
PubMed1997

Genes in idiopathic calcium oxalate stone disease.

An examination of the urinary excretions of 101 normal subjects indicated that the major genetic influence on calcium excretion is a codominant pair of alleles giving rise to three phenotypes, low, intermediate and high (hypercalciuric) excretors. This inference was based on variance, Hardy-Weinberg and segregation analyses. Similar independent gene pairs also appear to influence oxalate and citrate excretion, A 3-locus Hardy-Weinberg table using estimates of gene frequencies derived from the study of normals suggests that only 3 or 4 leading genes are involved in oxalate stone disease. Strong candidate genes identified from molecular and physiological studies cannot be proposed at present, but it is assumed that they influence the transport of these ions in either the intestine, kidney or both organs. The identification of the genes involved should be facilitated by the reduction of dietary influences on urinary excretions through the use of formula diets.

Calcium Oxalate↗
PubMed1997

The effects of (L)-2-oxothiazolidine-4-carboxylate on urinary oxalate excretion.

PURPOSE: A phase I study was done to evaluate the safety and pharmacokinetics of (L)-2-oxothiazolidine-4-carboxylate (OTZ). An ancillary objective was to compare the effects of treatment with 2 levels of OTZ to placebo on urinary oxalate excretion in healthy male subjects. MATERIALS AND METHODS: Individuals underwent intravenous infusion of 70 (6) or 100 (6) mg/kg, body weight OTZ, or placebo for 2 hours at 4, 8-hour intervals. Urine was collected during the 12 hours before treatment, and at 0 to 4, 4 to 8, 8 to 24, 24 to 28, 28 to 32 and 32 to 48 hours after the initial infusion. Urine samples were assayed for creatinine, oxalate, citrate, sulfate, urate, phosphate and pH. RESULTS: Urinary oxalate excretion relative to creatinine decreased significantly in the 100 mg./kg. dose group by 4.1 mg./gm. during the first 24 hours and by 4.6 mg./gm. in 24 to 48 hours compared to baseline values (p < 0.05). Slight decreases of 0.9 and 1.1 mg./gm., respectively, in the 70 mg./kg. dose group, and 1.6 and 2.3 mg./gm., respectively, in the placebo group were observed. Oxalate excretion on day 2 in the 100 mg./kg. dose group was significantly less than that in the placebo group (p = 0.04). Urinary pH decreased and sulfate excretion increased with OTZ therapy. CONCLUSIONS: Treatment with 100 mg./kg. OTZ every 8 hours decreases urinary oxalate excretion in healthy men.

Adult↗
PubMed1997

Pathways of hepatic oxalate synthesis and their regulation.

Important features of hepatic oxalate synthesis remain uncertain despite its clinical significance. To clarify the terminal steps of the biosynthetic pathway and their modulation, we have examined oxalate and glyoxylate synthesis in vitro using isolated guinea pig peroxisomes and purified lactate dehydrogenase (LDH). Glycolate was rapidly oxidized to glyoxylate by isolated peroxisomes followed by a slower conversion of glyoxylate to oxalate. The glycolate oxidase (GO)-catalyzed conversion of glyoxylate to oxalate was strongly inhibited by physiological concentrations of glycolate and lactate. In contrast, the LDH-catalyzed conversion of glyoxylate to oxalate was only marginally affected by physiological concentrations of lactate and unaffected by physiological glycolate concentrations. This inhibition pattern suggests that LDH, not GO, catalyzes this conversion in vivo. Alanine inhibited oxalate synthesis by converting the bulk of the glyoxylate to glycine. On exposure to high alanine concentrations, however, inhibition was not complete and peroxisomes were able to convert sufficient glycolate to oxalate to account for daily endogenous oxalate production. NADH was a potent inhibitor of oxalate production by LDH by increasing glycolate formation from glyoxylate. Glycine was an ineffective source of glyoxylate, and an alkaline pH, a high-glycine concentration, and a prolonged incubation time were required to obtain a detectable synthesis. These results suggest that oxalate synthesis will be modulated by the metabolic state of the liver and resultant changes in NADH, lactate, and alanine levels.

Alanine↗
PubMed1996

Intra-uterine growth retardation.

Small-for-gestational-age fetuses are frequently detected in general obstetric practice. Despite remarkable improvements in our understanding and management of this group of conditions over the past 20 years, much more research is required. During the year reviewed in this article, advances were reported in identification, association/causation, relationship to maternal smoking, fetal endocrinology, clinical investigation and management, and these are discussed in turn.

Delivery, Obstetric↗
PubMed1995

Glucagon increases urinary oxalate excretion in the guinea pig.

Factors that influence hepatic oxalate synthesis are poorly defined. Hormones are important regulators of hepatic metabolism and could potentially be involved. The effects of hyperglucagonemia were examined in guinea pigs injected with either saline or pharmacological doses of glucagon for 4 days. Glucagon treatment increased mean urinary oxalate excretion by 77% in male and 34% in female animals. The levels of hepatic peroxisomal enzymes involved in oxalate synthesis declined with glucagon treatment, but experiments with isolated peroxisomes indicated that oxalate synthesis in vitro was unaffected. Glucagon decreased hepatic alanine levels by 66%, lactate by 69%, and pyruvate by 73%, but glycolate and glyoxylate levels were unaffected. This decrease in alanine would substantially lower the activity of alanine-to-glyoxylate aminotransferase activity in vivo and make more glyoxylate available for oxalate synthesis. The decrease in lactate and pyruvate concentrations would stimulate the enzymatic conversion of glyoxylate to oxalate and may account for the increase in oxalate synthesis without an increase in glyoxylate concentration. These results demonstrate that hepatic oxalate synthesis is influenced by metabolic changes and that alterations in hepatic alanine, lactate, and pyruvate concentrations may be important elements.

Animals↗
PubMed1995

Measurement of urinary oxalate and citrate by capillary electrophoresis and indirect ultraviolet absorbance.

We describe a method for measuring urinary oxalate and citrate with capillary electrophoresis (CE) and indirect ultraviolet absorbance detection. Sample preparation is minimal, requiring an acidification, brief centrifugation, and dilution. The method is rapid, with oxalate and citrate having mean migration times of 4.02 and 4.50 min, respectively. The minimal detectable concentration (signal-to-noise ratio of 7) of both oxalate and citrate in urine was 7 mg/L. Total imprecisions (CV) were 1.2-5.6% for three urine samples with oxalate and citrate concentrations of 8-60 mg/L and 80-860 mg/L, respectively. The recovery of added oxalate ranged from 94% to 101%. Results of CE analyses agreed well with enzymatic determinations of oxalate and citrate. Rapid analysis time, accuracy, and reproducibility make this procedure well suited for routine urinary oxalate and citrate determinations.

Capillary Action↗
PubMed1995

Dietary oxalate and its intestinal absorption.

Dietary oxalate is currently believed to make only a minor contribution (< 20%) to urinary oxalate excretion. A recent prospective study of stone disease suggested that dietary oxalate may be a significant risk factor. This observation led us to re-evaluate the contribution of dietary oxalate to urinary oxalate excretion. Previous studies have been hampered by inaccurate food composition tables for oxalate and inadequate methods for studying intestinal oxalate absorption. This evidence as well as factors that modify oxalate absorption are reviewed. New approaches to measure food oxalate and intestinal oxalate absorption have been examined. Capillary electrophoresis appears to be well suited for the analysis of the oxalate content of food. Two individuals consumed an oxalate-free formula diet for 7 days. This diet decreased urinary oxalate excretion by an average of 67% (18.6 mg per 24 hours) compared to oxalate excretion on self-selected diets. The absence of detectable oxalate in feces by day 6 of the diet suggested that the intestinal absorption was minimal. However, an effect of the formula diet on endogenous oxalate synthesis cannot be excluded. Restoring oxalate to the formula diet increased urinary oxalate excretion and illustrates that this experimental protocol may be well-suited for studying oxalate absorption and factors that modify it. Our results suggest that the intestinal absorption of dietary oxalate makes a substantial contribution to urinary oxalate excretion and that this absorption can be modified by decreasing oxalate intake or increasing the intakes of calcium, magnesium, and fiber.

Animals↗