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Utilization of galactose in cultured brain cells of neonatal mice.

Metabolism of galactose was examined in dissociated brain cells from neonatal mice after 10-13 days in culture. Consumption of galactose at levels up to 26 mM was much less than consumption of glucose at corresponding concentrations. Lactate was consumed from the media at all galactose levels, in contrast to experiments with glucose in which lactate was formed and released into the media. Generation of CO2 from 4 mM glucose was 9-fold greater than from an equimolar level of galactose. Relatively low concentrations of glucose could reduce uptake of galactose, whereas galactose at levels up to 11.6 mM failed to inhibit consumption of glucose or formation of lactate. In glucose-deficient states, galactose supplementation of the media led to a marked increase in sulfatide synthesis by oligodendrocytes in the culture with a maximum effect at 2.3 mM. Under these conditions, [1-14C]galactose was incorporated directly into the carbohydrate portion of sulfatide, although most of the label was found in phospholipids and in the nonlipid fraction of the cellular homogenate. These data suggest that galactose is poorly metabolized by brain cells, but does not exhibit toxic effects.

Animals↗

Galactose assimilation in pups of diabetic canine mothers.

The effects of enteric galactose alimentation on neonatal glucose turnover and hepatic glycogen synthesis were investigated in a newborn animal model of diabetic pregnancy. Control pups and pups of diabetic dogs were studied in the basal state and after each group of pups was randomly fed equivalent amounts of galactose or glucose by oral-gastric tubes. Basal fasting blood glucose levels were not statistically different between the groups, whereas basal plasma insulin levels were 2-3 times higher in pups born to diabetic mothers. Blood glucose levels at each time point in response to glucose or galactose feeding in pups of diabetic mothers were not statistically different; however, the rise of plasma insulin concentrations was attenuated in pups of diabetic mothers fed galactose. The increase in the systemic rate of appearance of glucose and in glucose clearance were attenuated in pups of diabetic mothers fed galactose compared with those fed glucose. Hepatic glycogen content was augmented above basal levels in pups of diabetic mothers. Although glycogen synthase activity was not different between glucose- or galactose-fed pups of diabetic mothers, the active component of glycogen phosphorylase was reduced by both glucose and galactose feedings. Galactose alimentation had a greater effect on glycogen phosphorylase than did glucose alimentation. The observed increase in glycogen synthesis and reduced systemic glucose appearance after galactose alimentation could not be accounted for by the previously proposed excess of galactokinase over glucokinase activities when the latter enzyme was assayed at saturation. Indeed, neonatal hepatic glucokinase activity appeared to be induced during diabetic pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Endothelial changes in galactose-fed dogs.

Specular microscopic studies indicate that the size (polymegathism) and shape (pleomorphism) of the hexagonal corneal endothelial cells change in diabetics. Similar morphometric changes of the corneal endothelium have also been experimentally observed in diabetic rats as well as in diabetic and galactose-fed dogs and concomitant administration of aldose reductase inhibitors reduced these morphological changes. The purpose of this study was to examine whether corneal endothelial changes in galactose-fed dogs are reversible by the marked reduction of galactitol production after stopping prolonged galactose feeding. Ten control dogs were fed a normal diet, while 48 dogs were fed a diet containing 30% galactose. The galactose diet was removed from 15 dogs after 24 months at which time pericyte ghosts in the retina had developed and another 15 dogs were removed from the galactose diet after 31 months when retinal microaneurysms had developed. Eighteen dogs remained on galactose diet throughout the study (38 months). Specular microscopy was conducted on members of all groups after 38 months of study and the photographs were analyzed in masked fashion on the Bambi image analysis system. The evaluation of the corneal endothelial cells revealed significant differences in the cell size and density between galactose-fed dogs in the three groups and normal, age-matched control dogs. Corneal endothelial changes were not significantly reduced in dogs fed galactose for either 24 months or 31 months and then fed a normal diet for 14 and 7 months, respectively, indicating that amelioration of endothelial cell changes requires therapy prior to the advent of endothelial morphologic changes.

Animals↗

Galactose-conjugated antibodies in cancer therapy: properties and principles of action.

Galactose conjugation of antibodies causes them to be recognized by the hepatic asialoglycoprotein receptor and therefore cleared very rapidly from the blood. In these investigations, some effector functions of galactose-conjugated antibodies were assayed, and several applications to experimental tumors in vivo were demonstrated. Galactose conjugation did not interfere with two antibody functions in addition to antigen binding, namely complement-mediated cytotoxicity and antibody-dependent cell-mediated cytotoxicity. This conjugation procedure was originally developed for its potential use in localized immunotherapy, such as i.p. Injection of galactose-antibody conjugates i.p. demonstrated, more conclusively than other methods that have been used, that the presence of ascites causes prolonged retention of antibody in the peritoneal cavity and that this effect is correlated with the volume of ascites present. In mice bearing i.p. tumor xenografts, i.p. injection of galactose-antibody conjugates resulted in high tumor/nontumor ratios at 28 h after antibody injection, with values of 40:1, 43:1, 77:1, and 11:1 for the blood, kidney, lung, and spleen, respectively, although the ratio was only 4:1 for the liver. Control experiments demonstrated that i.p. injection of unconjugated antibody or a galactose-conjugated nonreactive antibody produced much lower tumor/nontumor ratios. In investigations of possible systemic application of galactose-antibody conjugates, we found that injection of large amounts of an inhibitor that binds competitively to the hepatic receptor, asialo-bovine submaxillary mucin, can block clearance of galactose-conjugated antibodies for 2-3 days. In this way, high blood levels of antibody can be maintained for 2-3 days, thus allowing penetration and binding to solid tumors, followed by very rapid blood clearance. With this approach, using a human carcinoma growing s.c. in nude mice, high tumor/nontumor ratios were obtained 4 days after injection, with mean values of 43:1, 18:1, 17:1, and 15:1 for the blood, kidney, lung, and spleen, respectively, although the ratio for the liver was only 1.7:1. The blood level at this time was 0.04 +/- 0.02% (SD) of the injected dose/g, while the tumor level was 1.69 +/- 1.29% of the injected dose/g. In conclusion, galactose-conjugated antibodies appear to have diverse applications in regional or systemic immunotherapy.

Animals↗

Stability of galactose in aqueous solutions.

The stability of 5%-30% w/v galactose in sterile water for injection and acetate and phosphate buffers was studied. The concentration of galactose was determined after each sample was diluted to a nominal concentration of 0.5% (w/v); for purposes of data analysis, the concentration as measured in the diluted sample was multiplied by a dilution factor to obtain the true concentration in the sample. The concentrations were determined from the regression line obtained by plotting the peak-height ratios (for various concentrations of galactose and the internal standard cellobiose) versus the galactose concentrations. Triplicate samples were quantitatively analyzed for galactose content by high-performance liquid chromatography. The stability of the samples was then studied in relation to buffer concentration; pH; storage at 25, 45, and 65 degrees C for six weeks, and autoclaving at 121 degrees C for 30 minutes. Galactose degradation increased in relation to its concentration, increasing temperature, and buffer concentration. Galactose solutions in water and phosphate incurred less than 5% degradation on autoclaving; however, the 30% solutions in acetate buffers lost up to 21% of initial content. Yellow discoloration of solutions was associated with autoclaving and prolonged exposure at 65 degrees C and appeared in some solutions that did not exceed the USP XXI limit of 5-hydroxymethylfurfural and related compounds in dextrose injection. The estimated room temperature shelf-life of galactose in sterile water for injection sterilized by 0.45-micron-porosity membrane filtration is four and one-half months. Solutions may also be sterilized by autoclaving at 121 degrees C for 30 minutes; galactose solutions containing pH buffers should not be sterilized by autoclaving.

Chromatography, High Pressure Liquid↗

Impaired oxidation of carbon-labeled galactose by alcoholic or diabetic liver in vivo.

Because of the organ and enzyme specificity of the metabolism of galactose, evaluation of various kinds of liver disease can be done by measuring the formation of labeled breath CO2 from carbon-labeled galactose in vivo. As shown earlier with uniformly 14C- or 13C-labeled galactose, a further study of alcoholic cirrhotic patients and controls with cheaper 1-14C-galactose indicates a superior discriminatory value of this test compared with common liver function tests. The oxidation test is easier to perform and more acceptable to patients than the standard galactose tolerance blood test. Output of 14CO2 showed slight correlations with serum albumin and 99mTc-sulfur colloid scan grade, but not with other function tests (SGOT, alkaline phosphatase, bilirubin). Comparison with five-year clinical outcome (two groups: with or without known liver-related death) in 29 of 43 total cirrhotic patients (U-14C or 1-14C-galactose) showed a low (75% probability) significance of prognosis for the galactose oxidation test, but none for any of the other tests. A two-part test of oxidation of 14C-galactose (with and without an acute dose of ethanol) in 19 possibly or likely alcoholic (but non-cirrhotic) persons indicated, by correlation with other liver function tests and drinking history, some possibly enhanced sensitivity of the two-part versus the single test for recognizing early liver damage. A preliminary study of the single galactose oxidation test in 7 patients with Type II diabetes suggests moderate impairment of oxidation, which might be applied to evaluate the hepatic disorder in diabetes.

Adult↗

Re-examination of the products of the action of galactose oxidase. Evidence for the conversion of raffinose to 6''-carboxyraffinose.

Galactose oxidase is a fungal enzyme which is known to oxidize the C-6 hydroxymethyl of galactose to an aldehyde group. When the products of a galactose oxidase-catalase treatment of raffinose were examined by gel filtration and ion exchange chromatography, we found that, in addition to the expected 6''-aldehydoraffinose, two other components were present. Of these two components, the major one was retained on a column of AG 1-X8 (formate), gave a positive carbazole reaction for uronic acid, and on paper chromatograms had a mobility identical with that of 6''-carboxyraffinose. The infrared spectrum of the compound showed a carbonyl absorbance at 1725 cm-1 and was distinguishable from the spectra of raffinose and 6''-aldehydoraffinose. These data showed that raffinose was partly converted to 6''-carboxyraffinose when treated with galactose oxidase and catalase. The conversion of [3H]raffinose to [3H]6''-carboxyraffinose increased gradually with time of oxidation from 22% at 6 h to 68% at 96 h. Results of other experiments provided evidence that this was an enzymic conversion and depended on the presence of galactose oxidase. The activities responsible for the formation of aldehyde and uronic acid could not be separated by affinity chromatography, gel electrophoresis, or ion exchange chromatography, indicating that the same enzyme is responsible for both activities. Treatment of galactose, melibiose, and stachyose with galactose oxidase and catalase also resulted in the formation of the corresponding uronic acids. These studies indicate that galactose oxidase not only converts the C-6 hydroxymethyl group of galactose to an aldehyde group, but also catalyzes further oxidation to the carboxyl group.

Animals↗

Bedside micro-method for measuring effective hepatic blood flow, with use of first-order galactose clearance pharmacokinetics.

Data from first-order galactose clearance were used to estimate "effective" hepatic blood flow in normal subjects and in patients with hepatic diseases. Galactose clearance was determined by infusing galactose intravenously at a constant rate and measuring its resulting steady-state concentration in blood. The infusion rate must not exceed the maximum rate of galactose clearance by the liver, the "galactose elimination capacity." With this constraint and within the physiological and pathophysiological limits of hepatic blood flow, a constant infusion at 50 mg/min results in steady-state concentrations ranging from 20 to 200 mg/L. To measure galactose within this range, we modified a YSI Model 23A glucose analyzer and, using an immobilized galactose oxidase (EC 1.1.3.9)/hydrogen peroxide electrode system, accurately measured galactose in water and blood. The galactose metabolic clearance rate was calculated for six normal rats and for a normal and a cirrhotic human subject. The speed of the analysis (40 s), the small sample required (25 microL), and the suitability of fresh whole blood as the sample make the method ideal for measuring hepatic blood flow in small laboratory animals as well as for determining at bedside the effective hepatic blood flow in humans.

Animals↗

Efficacy of treatment after measurable diabeticlike retinopathy in galactose-fed rats.

PURPOSE: To determine whether the diabeticlike retinal microangiopathies of the galactose-fed rat model could be ameliorated if intervention by withdrawal of the galactose diet or treatment with the aldose reductase inhibitor AL-3152 was initiated after quantifiable microangiopathies had occurred. METHODS: Weanling male Sprague-Dawley rats were randomized into five groups and fed for up to 24 months Purina laboratory chow (#5001) plus 50% starch (control [CON]), 50% D-galactose (galactose [GAL]), 50% D-galactose with AL-3152 (approximately 14 mg/kg per day) (prevention [PRV]), 50% D-galactose for 6 months followed by intervention with the inhibitor (intervention [INT]), or 50% D-galactose for 6 months followed by replacement with the 50% starch diet (withdrawal [GWD]). In rats on experimental diets and killed after 6, 18, and 24 months, one retina was prepared for transmission electron microscopy; the other was used for vessel wholemounts using elastase digestion. Capillary images were analyzed by computer morphometry. RESULTS: At 6 months, the GAL rats exhibited statistically significant (P < 0.05) increases over CON rats in mean capillary basement membrane thickness, capillary density, and dilated channels. These parameters tended to increase with time in most groups, and the differences between GAL and age-matched CON rats were maintained at the 18- and 24-month endpoints. Although the microangiopathies were ameliorated by AL-3152 treatment from the onset (PRV), intervention after 6 months of galactosemia with either galactose withdrawal (GWD) or addition of inhibitor (INT) showed amelioration in only some parameters at 18 months and no statistically significant benefit at the 24-month endpoint. CONCLUSIONS: Amelioration of galactose-induced retinal microangiopathies with AL-3152 in the prevention group suggests an efficacious application of aldose reductase inhibitors in treating diabetic retinopathy, provided treatment can begin soon after the onset of diabetes. Intervention after some of the earliest microscopic lesions neither halted progression of the angiopathy nor provided appreciable benefit at the 24-month follow-up.

Aldehyde Reductase↗

Characteristics of a Galactose-adapted Sugarcane Cell Line Grown in Suspension Culture.

Although d-galactose is normally toxic to sugarcane (Saccharum sp.) cells, a cell line that grows on 100 mm galactose has been propagated. Nonadapted cells in a medium containing galactose instead of sucrose accumulate UDP-galactose; these cells also have much lower UDP-galactose 4-epimerase (EC 5.1.3.2) activity than do adapted cells. This enzyme may determine whether or not galactose will cause toxicity symptoms to develop. The growth rate of galactose-adapted cells is similar to most cell lines on several other carbohydrates. The galactose-adapted cells are also similar to sucrose stock cells in cell wall composition and sugar phosphate concentrations, but, like the nonadapted cells, accumulate free galactose.

Journal Article↗

Selection of Galactose-Fermenting Streptococcus thermophilus in Lactose-Limited Chemostat Cultures.

Stock cultures of Streptococcus thermophilus are essentially galactose negative (Gal). Although both galactose 1-phosphate uridyl transferase and uridine-5-diphospho-glucose 4-epimerase are present, suggesting that the genes for the Leloir pathway exist, cells cannot induce high levels of galactokinase. Therefore, galactose is largely excreted when cultures are grown on lactose, and most strains cannot be readily adapted to grow on free galactose. Gal cultures were grown in a chemostat under lactose limitation in which high concentrations of residual galactose were present. Under this selection pressure, Gal organisms eventually took over the culture with all four strains examined. Gal cells had induced galactokinase, and three of the four strains grew on free galactose with doubling times of 40 to 50 min. When Gal organisms were grown on lactose in batch culture, the galactose moiety was only partially utilized while lactose was still present. As lactose was exhausted, and catabolite repression was lifted, the Leloir pathway enzymes (especially galactokinase) were induced and the residual galactose fermented. Neither phospho-beta-galactosidase activity nor the enzymes of the d-tagatose 6-phosphate pathway were detected in S. thermophilus. In contrast to Streptococcus cremoris and Streptococcus lactis, fermentation was homolactic with galactose in batch cultures and with lactose limitation in the chemostat. When mixed Gal-Gal cultures were repeatedly transferred in milk, the Gal cells became the dominant cell type. The Gal phenotype of stock cultures probably reflects their prolonged maintenance in milk.

Journal Article↗

The DeLey-Doudoroff Pathway of Galactose Metabolism in Azotobacter vinelandii.

Azotobacter vinelandii cell extracts reduced NAD and oxidized d-galactose to galactonate that subsequently was converted to 2-keto-3-deoxy-galactonate. Further metabolism of 2-keto-3-deoxy-galactonate required the presence of ATP and resulted in the formation of pyruvate and glyceraldehyde 3-P. Radiorespirometry indicated a preferential release of CO(2) at the first carbon position of the d-galactose molecule. This suggested that Azotobacter vinelandii metabolizes d-galactose via the DeLey-Doudoroff pathway. The first enzyme of this pathway, d-galactose dehydrogenase, was partially characterized. It has a molecular weight of about 74,000 Da and an isoelectric point of 6.15. The pH optimum of the galactose dehydrogenase was about 9. The apparent K(m)s for NAD and d-galactose were 0.125 and 0.56 mM, respectively. Besides d-galactose, the active fraction of this galactose dehydrogenase also oxidized l-arabinose effectively. The electron acceptor for d-galactose or l-arabinose oxidation, NAD, could not be replaced by NADP. These substrate specificities were different from those reported in Pseudomonas saccharophila, Pseudomonas fluorescens, and Rhizobium meliloti.

Journal Article↗

Carbohydrate specificity of the galactose-recognizing receptor of rat peritoneal macrophages.

The galactose-recognizing system of rat peritoneal macrophages mediates the binding and uptake of desialylated blood cells and glycoproteins. To characterize the specificity of this receptor, binding studies were performed with various galactose derivatives as competitive inhibitors and sialidase-treated erythrocytes or asialoorosomucoid as ligands for receptors, which were either membrane-bound or isolated after solubilization. From the results obtained it can be concluded that galactose is recognized via its hydrophobic and/or hydrophilic regions, formed by the accumulation of OH-functions on one side and of H-atoms on the other ("side effect"), whereas the binding partner or the anomeric configuration of galactose has no significant influence. Although it became apparent that not a single hydroxyl group of the sugar is responsible for binding, the hydroxyl at C-4 seems to be most important, followed by the OH-group at C-3. Those at C-1, C-2 and C-6 do not play a great role. This order of importance ("position effect") was found with galactose, derivatized by methylation or otherwise, and with diastereomers of galactose. Whereas the recognition of a single galactose residue leads to weak binding only, an appropriate arrangement of several of these ligands in one molecule results in an enormous increase in the binding strength of each galactose residue. This "cluster effect" was observed not only with membrane-bound but also with solubilized receptor. However, the binding of asialoorosomucoid by the latter was better inhibited with free galactose, when compared with the membrane-bound receptor.

Animals↗

Characterization of UDP-galactose:2-acetamido-2-deoxy-D-glucose 3 beta-galactosyltransferase from pig trachea.

We have previously reported the enzymatic synthesis of galactosyl-beta 1,3-N-acetylglucosamine using membrane preparations from pig trachea (Sheares, B. T., Lau, J. T. Y., and Carlson, D. M. (1982) J. Biol. Chem. 257, 599-602). The enzyme catalyzing the synthesis of this disaccharide, UDP-galactose:N-acetylglucosamine 3 beta-galactosyltransferase, has been solubilized and contaminating galactosyltransferases, including the UDP-galactose:N-acetylglucosamine 4 beta-galactosyltransferase and the UDP-galactose:N-acetylgalactosamine-mucin 3 beta-galactosyltransferase, were removed by affinity chromatography on alpha-lactalbumin-agarose, N-acetylglucosamine-agarose, and asialo-ovine submaxillary mucin bound to DEAE-Sephacel. UDP-galactose:N-acetylglucosamine 3 beta-galactosyltransferase and a yet unidentified UDP-galactose:N-acetylglucosamine 4 beta-galactosyltransferase, both not retained by the alpha-lactalbumin-agarose, were further purified by adsorption to a UDP-hexanolamine-Sepharose column and these two galactosyltransferase activities were finally resolved by gel filtration on Sephacryl S-200. The purified UDP-galactose:N-acetylglucosamine 3 beta-galactosyltransferase displays an absolute requirement for a divalent cation (Mn2+ or Co2+) and is most active at pH 6.0. Unlike the UDP-galactose:N-acetylglucosamine 4 beta-galactosyltransferase, UDP-galactose:N-acetylglucosamine 3 beta-galactosyltransferase is not inhibited by high concentrations of N-acetylglucosamine. Apparent Km values are UDP-galactose, 0.23 mM, and N-acetylglucosamine, 385 mM. The apparent Km for a synthetic acceptor, N-acetylglucosaminyl-beta 1, 3-N-acetylgalactosamine-O-benzyl, was 2.4 mM. Acceptor specificity suggests that this 3 beta-galactosyltransferase is responsible for elongation of oligosaccharide chains in mucin glycoproteins and in glycolipids.

Animals↗

Further observations in a case of uridine diphosphate galactose-4-epimerase deficiency with a severe clinical presentation.

The red-cell concentrations of galactose-1-phosphate and uridine diphosphate galactose have been studied in relation to dietary galactose in a case of uridine diphosphate galactose-4-epimerase deficiency (McKusick 23035). Uridine diphosphate galactose accumulates rapidly in response to very small amounts of galactose but the concentration of galactose-1-phosphate increases proportionately to galactose intake. The significance of the observation is discussed with respect to the pathogenesis and treatment of the disease.

Biotransformation↗

The binding of decomposition products of UDP-galactose to the microsomes and polyribosomes isolated from rat liver.

UDP-D-[U-14C]galactose is decomposed to [U-14C]galactose-1-phosphate and [U-14C]galactose by rat liver microsomal and crude polyribosomal fractions, under conditions commonly used to assay of glycosyltransferase activities. UDP-D-[U-14C]galactose, at neutral pH, is also chemically degraded to the [U-14C]galactose-1,2-cyclic phosphate. The 1,2-cyclic phosphate derivative of galactose also exists in the commercial UDP-D-[U-14C]galactose. It is a very important finding that products of the UDP-D-[U-14C]galactose decomposition are tightly, although nonenzymatically, bound to tested subcellular fractions and may create a false impression of protein glycosylation. The application of controls containing all radioactive substances present in suitable samples is recommended in order to avoid incorrect interpretations of the results.

Animals↗

Transport of UDP-galactose in plants. Identification and functional characterization of AtUTr1, an Arabidopsis thaliana UDP-galactos/UDP-glucose transporter.

The synthesis of non-cellulosic polysaccharides and glycoproteins in the plant cell Golgi apparatus requires UDP-galactose as substrate. The topology of these reactions is not known, although the orientation of a plant galactosyltransferase involved in the biosynthesis of galactomannans in fenugreek is consistent with a requirement for UDP-galactose in the lumen of the Golgi cisternae. Here we provide evidence that sealed, right-side-out Golgi vesicles isolated from pea stems transport UDP-galactose into their lumen and transfer galactose, likely to polysaccharides and other acceptors. In addition, we identified and cloned AtUTr1, a gene from Arabidopsis thaliana that encodes a multitransmembrane hydrophobic protein similar to nucleotide sugar transporters. Northern analysis showed that AtUTr1 is indeed expressed in Arabidopsis. AtUTr1 is able to complement the phenotype of MDCK ricin-resistant cells; a mammalian cell line deficient in transport of UDP-galactose into the Golgi. In vitro assays using a Golgi-enriched vesicle fraction obtained from Saccharomyces cerevisiae expressing AtUTr1-MycHis is able to transport UDP-galactose but also UDP-glucose. AtUTr1- MycHis does not transport GDP-mannose, GDP-fucose, CMP-sialic acid, UDP-glucuronic acid, or UDP-xylose when expressed in S. cerevisiae. AtUTr1 is the first transporter described that is able to transport UDP-galactose and UDP-glucose. Thus AtUTr1 may play an important role in the synthesis of glycoconjugates in Arabidopsis that contain galactose and glucose.

Amino Acid Sequence↗

Sugar nucleotide concentrations in red blood cells of patients on protein- and lactose-limited diets: effect of galactose supplementation.

Uridine diphosphate (UDP) galactose, a pivotal compound in the metabolism of galactose, is the obligate donor of galactose in the formation of complex glycoconjugates. The cellular UDPgalactose concentration has been thought to be maintained by the interconversion of UDPglucose and UDPgalactose by UDPgalactose-4-epimerase. However, recent findings of lower average red blood cell (RBC) UDPgalactose concentrations in galactose-1-phosphate uridyltransferase-deficient patients suggest that other factors play a role in determining its concentration. To test the hypothesis that the amount of galactose traversing the Leloir pathway contributes to the cellular UDPgalactose pool, we determined RBC UDPgalactose in patients with maple syrup urine disease (MSUD), phenylketonuria (PKU), and other metabolic diseases who were treated with a low-protein, and consequently, low-lactose diet. Six patients with MSUD were also supplemented with 19 g galactose/d and their UDPhexose concentrations were measured at intervals. We show that young patients with MSUD or PKU have decreased average RBC UDPgalactose concentrations when compared with similarly aged healthy subjects. Galactose supplementation of MSUD patients significantly increased their UDPgalactose concentrations in both RBCs and white blood cells (WBCs) from 29.5 +/- 1.5 to 42.3 +/- 5.8 nmol/g hemoglobin and from 69.0 +/- 7.5 to 193.0 +/- 49.0 nmol/g protein, respectively. Discontinuation of supplementation was associated with a return to basal values in RBCs and a reattainment of the pretreatment ratio of UDPglucose to UDPgalactose in WBCs. These observations demonstrate that dietary galactose is a factor in establishing the steady state concentrations of the uridine sugar nucleotides and imply that galactose metabolism modulates the achievement of an epimerase-mediated equilibrium.

Adolescent↗