Search PubMed⌕ Search

Biomedical subjects

S H Mudd

Publications and source records attributed to S H Mudd.

At least 73 records · Page 4Linked to original sources
PubMed1981

Homocysteine Biosynthesis in Green Plants: Physiological Importance of the Transsulfuration Pathway in Lemna paucicostata.

To permit an assessment of the relative contributions of the transsulfuration and the direct sulfhydration pathways for homocysteine biosynthesis, the time course of incorporation of (35)S from (35)SO(4) (2-) into various sulfur-containing compounds in Lemna paucicostata has been determined. Plants were grown with either low (4.5 micromolar) or ample (1,000 micromolar) sulfate in the medium. At the shortest labeling times, (35)S-cystathionine was the predominant (35)S-containing organic sulfur compound. The flux of sulfur into cystathionine was sufficient to sustain the known rate of methionine biosynthesis. It was calculated that transsulfuration accounted for at least 90 and 85% of the total homocysteine synthesis in low and ample sulfate-grown plants, respectively (and may have accounted for 100%). No marked rise in the (35)S-soluble cysteine:(35)S-homocysteine ratio was observed even at the shortest labeling times, but it is argued that this may be due to (a) the observed compartmentation of soluble cysteine, and (b) the impracticality of using labeling times shorter than 17 seconds. Additional evidence supporting the importance of transsulfuration in Lemna is briefly described.

Journal Article↗
PubMed1981

A study of cardiovascular risk in heterozygotes for homocystinuria.

Early atherosclerotic-like lesions and thromboemobolic problems are prominent in homocystinuric patients. Recent evidence suggested that mild homocyst(e)inemia, such as is present in heterozygotes for homocystinuria due to cystathionine synthase deficiency, may cause a marked excess in early ischemic heart disease. To evaluate the risk due to mild homocyst(e)inemia, the frequencies of heart attacks and strokes in parents and grandparents of homocystinuric children were assessed in the present study. No statistically significant increases in the incidence of heart attacks or strokes were consistently detected. The data available are sufficient to virtually exclude an increase in the cardiovascular risk for homocystinuria heterozygotes of as much as fivefold compared to controls, and to make very improbable a relative risk of as much as threefold. Less than 5% of homocystinuria heterozygotes are likely to have a fatal or nonfatal heart attack by age 50. These results fail to suggest that mild homecyst(e)inemia is an important contributory factor in the overall incidence of cardiovascular disease.

Adult↗
PubMed1980

Epileptiform ocular movements with methylmalonic aciduria and homocystinuria.

A 7 1/2-year-old girl with a rare defect in cobalamin (vitamin B12) metabolism ("cobalamin C" type) developed epileptiform ocular and eyelid movements as the major clinical manifestation of the disease. One of three other patients who have been described with congenital syndrome was similarly noted to have "fluttering" of the eyelids interpreted as epileptic discharges. The metabolic abnormality produced a defect in synthesis of cobalamin coenzymes. It is characterized biochemically by the excreation of methylmalonic acid and homocystine in the urine.

Child↗
PubMed1980

Lemna paucicostata Hegelm. 6746: DEVELOPMENT OF STANDARDIZED GROWTH CONDITIONS SUITABLE FOR BIOCHEMICAL EXPERIMENTATION.

Photoautotrophic and mixotrophic growth of Lemna paucicostata Hegelm. 6746 (formerly Lemna perpusilla Torr. 6746) was investigated to establish standardized conditions for biochemical studies. Optimal temperature for growth was 29 to 30 C. The medium used previously (Datko AH, Mudd SH, Giovanelli J 1977 J Biol Chem 252: 3436-3445) was modified by inclusion of NH(4)Cl, decreasing macronutrient and ethylenediamine tetraacetate concentration, increasing micronutrient concentration, and inclusion of bicarbonate (for photoautotrophic growth) or 2-(N-morpholino)ethanesulfonic acid (for mixotrophic growth) buffers. Varying the sulfate concentration between 14 and 1 millimolar had no effect on growth. For photoautotrophic growth in the new medium (medium 4), the effects of CO(2) concentration, light intensity, and pH were measured. Under the optimal conditions, a multiplication rate (MR) of 300 to 315, equivalent to a doubling time of 23 to 24 hours was obtained. Addition of glutamine or asparagine did not increase this MR. For mixotrophic growth in low light, the effects of sucrose concentration and pH were determined. Under optimal conditions, MR was 210. A concentration of sucrose less than maximal for growth was chosen for the medium for experiments which will include (14)C-labeling of intermediates. MR under these conditions was 184. Growth was equally good in medium 4 and in half-strength Hutner's medium when sulfate was high (0.4 to 1 millimolar), but better in medium 4 when sulfate was low (20 micromolar). Growth rates could be restored to normal in half-strength Hutner's with low sulfate by decreasing the molybdate concentration.By modifying medium 4 to contain very low amounts of sulfate, and by preconditioning medium and plants, it was shown that there was an increment in plant protein of approximately 2.5 micrograms per nanomole of added MgSO(4). Colonies undergoing sulfur limitation exhibited a slow growth rate and a high frond to colony ratio. Molybdate and selenate produced growth inhibition reversible by sulfate. Conditions were developed in which the plants could be maintained indefinitely in the presence of either molybdate or selenate in altered metabolic steady-states with lowered growth rates and protein per frond.

Journal Article↗
PubMed1980

Lemna paucicostata Hegelm. 6746: LIFE CYCLE AND CHARACTERIZATION OF THE COLONY TYPES IN A POPULATION.

The sequence of frond emergence and the intervals required for daughter colony separation have been determined for Lemna paucicostata Hegelm. 6746 growing under standardized conditions. After separation of a new mother colony, the first daughter colony is produced from the left meristematic pocket and separates after approximately 60 hours, the second daughter, produced from the right pocket, separates after a further 30 hours, and the third daughter, again from the left, after a further 40 hours. The pattern of alternating longer and shorter intervals for separation of daughters continues throughout the life of the mother colony.Protein contents of fronds and whole colonies were determined either by chemical methods or by labeling to isotopic equilibrium with (35)SO(4) (2-). The smallest fronds measured were 5 to 6% of the final area they would attain when fully expanded and contained 10% of the protein they would finally contain. Thus, most protein accumulation occurred during the phase of rapid frond expansion rather than in an earlier primordial stage. The specific rates of protein accumulation were 0.027 to 0.030 micrograms per microgram protein per hour during rapid frond expansion and relatively constant thereafter at 0.014 to 0.019 for whole colonies until at least four daughter colonies had separated.A substantial amount of (35)S is transferred directly from the mother frond to its attached daughter fronds. The results are consistent with the transfer of soluble compounds arising through turnover of protein in the mother frond.Tree diagrams and a mathematical treatment were utilized to relate the distribution of colony types and the over-all doubling time in a large population of Lemna colonies to the doubling times and modes of division of the individual colonies within that population. It was shown that a population with the growth properties described increases exponentially and that within a relatively few generations reaches an equilibrium population distribution which is independent of the initial proportions of the members of the population. The measured distribution of colony types producing odd-numbered as compared to those producing even-numbered daughters was the same as that predicted by the theoretical analyses. Once a mass culture of Lemna paucicostata has attained its equilibrium distribution of colony types, under our standard conditions the proportions of colonies in which the indicated daughter will be next to separate will be: 60%, first; 15%, second; 11%, third; 5%, fourth; 8% fifth or higher. Thus, about 91% of colonies will have produced no more than four daughter colonies. For at least this period the specific rate of protein accumulation is relatively constant and senescent changes in mother fronds are rare. In the equilibrium distribution, 39% of colonies will themselves be first daughters, 25% will be second daughters, 14% will be third daughters, 9% fourth, and 13% greater than fourth daughters. These combined results suggest that obtaining representative samples of colonies from such a population for biochemical studies should be relatively simple.

Journal Article↗
PubMed1979

Diseases of sulphur metabolism: implications for the methionine-homocysteine cycle, and vitamin responsiveness.

Sixteen inherited human diseases are now recognized, affecting most of the major steps in sulphur metabolism. Studies of patients with three types of homocystinuria have demonstrated unequivocally the major role of cystathionine formation in degradation of homocysteine, and the importance of homocysteine remethylation. Methionine balance studies of normal subjects and of a sarcosine oxidase-deficient subject have shown the predominant role of creatine synthesis in methionine utilization and permitted assessment of the rate of oxidation of the methyl group of methionine. Together, the results demonstrate that once regulatory adjustments have been made the rate of methylneogensis is nicely controlled so that labile methyl groups are made available in amounts just sufficient to meet the needs for methionine. When excess methionine is ingested the four-carbon moiety is diverted into cystathionine, the methyl group is oxidized via sarcosine and the flow of partially oxidized one-carbon units is diverted away from 5-methyltetrahydrofolate toward CO2. Studies of cystathionine synthase-deficient patients demonstrate that the capacity to respond or not to respond to pyridoxine administration is genetically controlled, probably through structural differences in mutant cystathionine synthases. However, the properties of the enzyme crucial in conferring responsiveness have not yet been identified.

Adolescent↗
PubMed1978

Homocysteine biosynthesis in green plants. Physiological importance of the transsulfuration pathway in Chlorella sorokiniana growing under steady state conditions with limiting sulfate.

The physiological roles of the transsulfuration and direct sulfhydration pathways in Chlorella sorokiniana growing under steady state photoautotrophic conditions with limiting sulfate were studied by following the patterns of assimilation of 35SO4(2-) into sulfur amino acids. The labeling patterns expected of each pathway were defined by means of models based on the rates of net synthesis of the terminal pools of GSH, protein cysteine, and protein methionine. The labeling patterns observed are entirely consistent with the transsulfuration pathway and inconsistent with the direct sulfhydration pathway. By analysis of the amounts of radioactivity present in key intermediates at labeling times as short as 1 s, it was demonstrated that direct sulfhydration makes no detectable contribution to homocysteine biosynthesis, and if operative contributes no more than approximately 3% of the total homocysteine biosynthesized. From the combined determinations of the initial rates of labeling and net rates of synthesis of the various sulfur amino acids, a tentative working model is presented that summarizes our best current estimates of the major fluxes of sulfur in the experimental system. The labeling data further showed that soluble cysteine consists of at least two pools. One pool, termed "rapidly turning over" cysteine comprises less than 1% of the total soluble cysteine, and is the precursor of GSH, protein cysteine, and, almost certainly, cystathionine. The other pool, "slowly turning over" cysteine, appears to be in equilibrium with "rapidly turning over" cysteine, but not to be further metabolized.

Chlorella↗
PubMed1978

Phytostat for the growth of lemna in semicontinuous culture with low sulfate.

An apparatus is described by means of which Lemna perpusilla 6746 was grown photoautotrophically at a series of constant concentrations of inorganic sulfate, as low as 0.26 mum. Theoretical considerations relevant to this system are discussed and examples of the operation of the apparatus are presented. The data obtained were used to calculate sulfate uptake by the plant colonies as a function of sulfate concentration. The apparatus should be useful for the production of relatively large quantities of Lemna, grown under highly uniform conditions, with or without limitation of one or more nutrient(s). Other small vegetatively reproducing aquatic plants could be similarly studied. Uptake studies could be carried out on a variety of plant materials.

Journal Article↗
PubMed1978

Sulfur-containing Compounds in Lemna perpusilla 6746 Grown at a Range of Sulfate Concentrations.

Lemna perpusilla 6746, grown photoautotrophically at a series of sulfate concentrations ranging from 0.32 to 1,000 mum, was labeled to radioisotopic equilibrium with (35)SO(4) (2-). Sulfur-containing compounds were isolated and purified from the colonies. Radioactivity in each compound was a measure of the amount of that compound present in the tissue. The following compounds were identified and quantitated: inorganic sulfate, glutathione, homocyst(e)ine, cyst(e)ine, methionine, S-methylmethionine sulfonium, S-adenosylmethionine, S-adenosylhomocysteine, cystathionine, chloroformsoluble (presumed to be sulfolipid), protein cyst(e)ine, and protein methionine. gamma-Glutamylcyst(e)ine, erythro- and threo-thiothreonine, and S-methylcysteine were not detected. No volatile (35)S compounds were formed during plant growth at 1,000 mum sulfate, nor were significant amounts of (35)S compounds excreted into the medium.The amount of each component present in colonies grown over the 3,000-fold range of medium sulfate was relatively constant except for inorganic sulfate. This increased about 30-fold from the lowest to the highest medium sulfate concentration. The total soluble sulfur amino acids increased about 1.5- to 2-fold, due primarily to an increased amount of glutathione. Protein cyst(e)ine and protein methionine were the major organic sulfur compounds in Lemna, and the amounts of these compounds remained virtually constant despite the variation in external sulfate concentration.Procedures for the analysis of S-adenosylmethionine, S-methylmethionine sulfonium, and S-adenosylhomocysteine are presented.

Journal Article↗
PubMed1976

Platelet survival and morphology in homocystinuria due to cystathionine synthase deficiency.

In homocystinuria due to cystathionine synthase deficiency thromboembolism is a major cause of mortality and morbidity. Recent studies by others identified an abnormally shortened platelet survival and increased platelet vacuolization in patients with homocystinuria. When we studied six additional patients, however, we found the platelet survival to be within normal limits for each. The mean survival (+/-1 S.D.) was 9.75+/-0.94 days (normal, 9.27+/-1.06). In addition, platelets from five patients with homocystinuria and three obligate heterozygotes could not be distinguished from those of seven normal control subjects by electron microscopy. Specifically, no increased vacuolization was observed. Genetic heterogeneity, technical differences of differences in plasma homocystine concentrations could account for these descrepant results. The mechanism of thrombosis in homocystinuria remains an open question.

Adult↗
PubMed1976

Cobalamins in fibroblasts cultured from normal control subjects and patients with methylmalonic aciduria.

The intracellular content and proportional distribution of B12 (cobalamin) derivatives in fibroblasts cultured from patients with various forms of methylmalonic aciduria, as well as from normal control subjects, has been determined by a two-dimensional chromatobioautographic technique. Each line of fibroblasts was grown in the presence of four concentrations of cobalamin, ranging from the 0.04-0.07 pmol/ml contained in the basal medium to 74 pmol/ml (100 ng/ml), added in form of hydroxocobalamin (OH-CHl). Control cells grown in the basal medium contained substantial proportions of both methylcobalamin (MeCbl) and adenysylcobalamin (AdoCbl), with the former predominating. As increasing concentrations of OH-CBl were added to the growth medium, the total cellular cobalamin content increased without marked changes in the relative proportions of MeCbl, AdoCbl, and OH-Cbl. Three different patterns were discernable in the cobalamin distributions of the cells cultured from patients with methylmalonic aciduria (Table 1 and Fig. 1).

Amino Acid Metabolism, Inborn Errors↗
PubMed1975

Folate-responsive homocystinuria and "schizophrenia". A defect in methylation due to deficient 5,10-methylenetetrahydrofolate reductase activity.

Homocystinuria and homocystinemia without hypermthioninemia, but with reccurent episodes of folate responseive schizophrenic-like behavior, was documented in a mildly retarded adolescent girl who lacked the habitus associated with cystathionine synthase deficiency. Enzymes involved in homocysteine-methionine metabolism were demonstrated to be normal. A defect in the ability to reducte N-5-10--methylenetetrahydrofolate to 5-methyltetrahydrofolate was demonstrated. Methylenetetrahydrofolate reductase was 18 per cent of control values. Methyltetrahydrofolate is used for the methylation of homocysteine to methionine, and a deficiency of this compound could explain the homocystinemia and homocystinuria.

Adolescent↗
PubMed1975

Cystathioninuria and homocystinuria.

Three circumstances prompted us to reexamine the relationship between abnormal cystathionine accumulation and possible homocystinuria resulting from this condition: (a) discovery of an infant girl with apparently alternating massive cystathioninuria and homocystinuria; (b) the presence of homocystinuria in some, but not all, previously reported cases of cystathioninuria probably due to gamma-cystathionine deficiency; and (c) the recent demonstration that mammalian cystathionine beta-synthase can cleave cystathionine to homocysteine. The following conclusions were reached: (a) Homocystine may arise as a result of bacterial contamination of a urine sample initially containing cystathionine, but not homocystine. (b) After a methionine load, a cystathioninuric patient may excrete readily detected amounts of homocystine. (c) However, homocystinuria is not a necessary concomitant of even massive cystathioninuria. These findings and some of their implications are briefly discussed.

Amino Acid Metabolism, Inborn Errors↗