Showing papers on "Thiamine published in 1978"

Thiamine deficiency Selective impairment of the cerebellar serotonergic system

Abstract: To explore the role of thiamine deficiency in synaptic transmission, the high-affinity uptake and release systems for putative neurotransmitters were studied in synaptosomal preparations isolated from the telencephalon, hypothalamus, and cerebellum of rats made thiamine deficient by diet or pyrithiamine. There was significant decrease in the uptake of serotonin by the synaptosomal preparations of the cerebellum. Although thiamine and its phosphorylated forms added in vitro did not restore the decreased serotonin uptake, the administration of the vitamin in vivo resulted in a significant reversibility of the inhibition of serotonin uptake, coinciding with dramatic clinical improvement. The study supports the possibility of an important serotonergic innervation of the cerebellum and suggests a selective involvement of this system in the pathogenesis of some of the neurologic manifestations of thiamine deficiency.

Human infections caused by thiamine- or menadione-requiring Staphylococcus aureus.

Abstract: Stable dwarf forms of Staphylococcus aureus have been identified in clinical specimens as the sole or predominant isolate in eight cases. These organisms have been shown to be menadione or thiamine dependent, i.e., cultivation in the presence of one of these agents has permitted growth of colonies which appear typical of S. aureus. In vitro resistance to aminoglycosides was overcome by cultivation in the presence of menadione or thiamine. Menadione- or thiamine-requiring S. aureus can be considered as causative agents in severe human infections. Special care must be taken if they are to be identified in pathological specimens. Their antibiotic sensitivity testing should be done comparatively on supplemented and nonsupplemented media.

Prevention of the Wernicke-Korsakoff syndrome: a cost-benefit analysis.

Abstract: The Wernicke-Korsakoff syndrome is a thiamine-deficiency disorder occurring primarily among alcoholics. To determine the economic feasibility of preventing this disease by fortification of alcoholic beverages with thiamine, we compared the cost of fortification with the cost of institutionalizing alcoholics with the disorder. The estimated annual incidence of institutionalization is eight per million adult population. The cost of long-term institutionalization, discounted to present value, is $70 million per year. The cost of adequately fortifying alcoholic beverages is estimated to range from $3 million per year if allithiamines are used, to as much as $17 million per year if thiamine hydrochloride proves necessary. Thus, the cost-benefit ratio may range from 1:23 to 1:4. It is economically advantageous to prevent the Wernicke-Korsakoff syndrome by fortification of alcoholic beverages with thiamine. The stability, safety and marketability of thiamine and the allithiamines in alcoholic beverages ...

Early edematous lesion of pyrithiamine induced acute thiamine deficient encephalopathy in the mouse

Abstract: Pyrithiamine induced acute thiamine deficient encephalopathy in the mouse is one of the possible animal models of human Wernicke-Korsakoff syndrome. In this experiment, the adult male Swiss Mice, treated with a daily subcutaneous injection of pyrithiamine in conjunction with a thiamine deficient diet, abruptly developed unique encephalopathic signs on day 10. In the animals sacrificed immediately after the onset of the disease, the gross examination of the brains revealed a small number of minute hemorrhagic lesions in the thalamus, mammilary bodies and pontine tegmentum, including the medial and lateral vestibular nuclei. When spared the hemorrhage, these regions appear intact grossly and in paraffin sections, but were found to be significantly altered in Epon sections. In semithin Epon sections of the pontine tegmentum, there was edematous swelling of all the astrocytes and oligodendrocytes and occasional myelin sheaths. By electron microscopy, the edema of astrocytes involved both nucleus and cytoplasm extensively. The oligodendroglial edema was severe in the peripheral cytoplasm, particularly in the inner loops of the myelin sheaths and only moderately in the nuclei, perinuclear cytoplasm and outer loops. Disintegration of the myelin lamellae occurred when edema of the inner loops had advanced. The axis cylinders surrounded by the edematous loops were essentially intact. In contrast to such glial cell damage, the nerve cells and blood vessels were not altered. These findings suggest that (1) astroglia and oligodendroglia are the cells most sensitive to thiamine deficiency and (2) the resultant glial cell injury is the initial change of thiamine deficient encephalopathy in man and in experimental animals.

The determination of thiamin pyrophosphate in blood and other tissues, and its correlation with erythrocyte transketolase activity

Abstract: A sensitive method for the specific measurement of thiamin pyrophosphate (TPP) has been developed using the apoenzyme recombination concept. Yeast pyruvic decarboxylase apoenzyme can be reconstituted by the addition of TPP or samples containing TPP, yielding the holoenzyme with activity proportionate to the amount of TPP added. Using this technique, reaction mixtures containing 0.2 to 1.5 ng TPP can be assayed. Normal human erythrocyte TPP ranges from 50 to 150 ng per ml packed cells. When rats are fed a thiamin deficient diet, the erythrocyte TPP level falls more rapidly than the erythrocyte transketolase activity. After 8 days, the level of TPP in the erythrocytes of deficient animals was 10% of the level in pair-fed controls. At this time, however, there was no appreciable decrease in their respective transketolase activities. The level of TPP in the liver also is decreased drastically after 8 days. Therefore it appears that erytyrocyte and liver TPP stores have begun to be depleted and suggest that erythrocyte TPP levels are a more sensitive indicator of thiamin status.

Detection and incidence of B and C vitamin deficiency in alcohol-related illness.

Abstract: The activity of the red blood cell enzymes transketolase, glutathione reductase, and aspartate transaminase, and their activation by the coenzymes thiamine, riboflavin, and pyridoxine, the pyruvate tolerance test, the leucocyte vitamin C concentration, and the activity in serum of gamma-glutamyl transferase were measured in a series of 35 patients with alcohol-related illness. The incidence of thiamine deficiency was 31% as assessed by the activation of transketolase, and 55% as assessed by the pyruvate tolerance test. The incidence of riboflavin deficiency was 23% and of ascorbic acid deficiency 91%. No cases of pyridoxine deficiency were detected. The pyruvate tolerance test was found to be a more sensitive test of thiamine deficiency than the transketolase activation, and the activation of red blood cell aspartate transaminase was found to be a poor indicator of pyridoxine deficiency. There was a poor correlation of the gamma-glutamyl transferase activity with the degree of vitamin deficiency, suggesting that alcohol exposure is only partly responsible for the observed vitamin deficiency.

The Aetiology of the Sudden Infant Death Syndrome: Current Ideas on Breathing and Sleep and Possible Links to Deranged Thiamine Neurochemistry

Abstract: Summary: The aetiology of the sudden infant death syndrome: Current ideas on breathing and sleep and possible links to deranged thiamine Neurochemistry. The sudden infant death syndrome (SIDS) is now the commonest cause of death between one week and one year of age in most western countries. An asphyxia/ death, with unrecognised hypoxic episodes during sleep in the preceding weeks, has been postulated from autopsy evidence for both acute and chronic hypoxia; the evidence includes Po2 values, intra-thoracic distribution of petechiae, pulmonary arteriolar and right ventricular hypertrophy. Long-term monitoring of infants resuscitated from a “near miss” SIDS demonstrates sleep apnoea, sometimes associated with episodic collapse and obstruction of the upper airway. Physiological studies in healthy babies and animals highlight factors leading to vulnerability to asphyxia in different phases of sleep. In REM-sleep (rapid-eye-movement), inhibition of intercostal muscle activity leads to: inspiratory collapse of the rib-cage, impaired reflex compensation for airway obstruction, overall lung-deflation with reduction of O2-stores and rapid hypoxaemia during apnoea. In REM-sleep, breathing efforts are not augmented by hypercapnia and the defense against asphyxia depends on reflex responses to hypoxia. Sleep apnoea sometimes occurs in infants with a rare congenital defect of brain thiamine triphosphate. This draws attention to many similarities of modern SIDS and other infantile syndromes reported historically which involve deranged thiamine neurochemistry. Sudden unexpected deaths occur in apparently thriving infants of asymptomatic thiamine deficient mothers. Other similarities include: a peak incidence at 2–4 months of age; precipitation often by minor febrile episodes; seasonal and familial risk factors, with increased risk in twins; many common findings at autopsy. Although asymptomatic maternal thiamine deficiency is common in western communities ingesting high carbohydrate diets containing various thiamine antagonists, the effect on infant thiamine stores has received little attention. Future research is needed to evaluate SIDS incidence after identification and elimination of low thiamine states. Defective neural control of breathing during sleep should be evaluated in relation to thiamine-neurochemistry, particularly to the leaky blood-brain barrier, to glutamate and GAB A, to sympathetic denervation and to defective vagal reflexes of the lungs and larynx.

Binding and transport of thiamine by Lactobacillus casei.

Abstract: The relationship between thiamine transport and a membrane-associated thiamine-binding activity has been investigated in Lactobacillus casei. Thiamine transport proceeds via a system whose general properties are typical of active uptake processes; entry of the vitamin into the cells requires energy, is temperature dependent, exhibits saturation kinetics, and is inhibited by substrate analogs. A considerable concentration gradient of unchanged thiamine can be achieved by the system, although the vitamin is slowly metabolized to thiamine pyrophosphate. Consistent with these results, L. casei also contains a high-affinity, thiamine-binding component which could be measured by incubation of intact cells with labeled substrate at 4 degrees C (conditions under which transport is negligible). Binding was insensitive to iodoacetate, occurred at a level (0.5 nmol per 10(10) cells) nearly 20-fold higher than could be accounted for by facilitated diffusion, and was found to reside in a component of the cell membrane. Participation of this binder in thiamine transport is supported by the observations that the processes of binding and transport showed similarities in their (i) regulation by the concentration of thiamine in the growth medium, (ii) binding affinities for thiamine, and (iii) susceptibility to inhibition by thiamine analogs.

Active transport of thiamine by freshly isolated rat hepatocytes.

Abstract: Rat hepatocytes were freshly prepared from adult animals using the collagenase-perfusion technique. The hepatic transport of thiamine was studied in isolated liver cells. The process was found to be saturable with an apparent Kt of 0.31mM and a Vmax of 0.7 μmoles/ml intracellular fluid/5 minutes. However, at higher substrate concentrations, the process proceeded in a linear fashion. Both pyrithiamine and oxythiamine were inhibitory on the hepatic uptake of thiamine, the latter showed much weaker activity than the former. The system required the presence of sodium ions and was sensitive to ouabain. Anaerobic condition and metabolic inhibitors, e.g., 2, 4-dinitrophenol, cyanide, and iodoacetate suppressed the uptake rate of thiamine. Addition of ethanol in the incubation medium also caused significant reduction of thiamine uptake. Efflux studies indicated that a portion of intracellular thiamine is readily available for exodus. Chromatographic analyses showed that thiamine was only slightly metabolically altered during the transport process. It is suggested that thiamine is transported into isolated hepatic cells by an active, sodium-dependent process.

In vivo and in vitro response of human branched chain alpha-ketoacid dehydrogenase to thiamine and thiamine pyrophosphate.

Abstract: Summary: In a homozygous affected patient with maple syrup urine disease, pharmacologic doses of thiamine lowered urinary excretion of branched chain α-ketoacids and stimulated branched chain α-ketoacid dehydrogenase (BCKAD) in his peripheral blood leukocytes. Supplementation of his branched chain aminoacid restricted diet with 100 mg/day of thiamine eliminated recurrent episodes of ketoacidosis. These clinical responses were studied in vitro using mitochondrial inner membranes prepared from his cultured skin fibroblasts and those from another thiamine-responsive patient from Canada. BCKAD in both mutant cell lines had similarities to normal enzyme including: identical apparent Km value for thiamine pyrophosphate; similar heat inactivation profiles which were slowed by the presence of thiamine pyrophosphate; and stimulation above basal activity by thiamine pyrophosphate. Differences in the enzymes included: decreased apparent Vmax for thiamine pyrophosphate; increased lability at 37°; and failure to respond to added NAD+ CoASH, and Mg2+. We propose that “excess” thiamine led to increased available thiamine pyrophosphate which stabilized the branched chain α-ketoacid dehydrogenase, decreased biologic turnover, increased enzyme specific activity and produced in vivo tolerance to branched chain aminoacids in these patients with maple syrup urine disease. Speculation: By studying the partially purified normal and mutant branched chain α-ketoacid dehydrogenases from cultured human fibroblasts, direct in vitro effects of thiamine pyrophosphate can be measured and related to in vivo clinical responses. This should improve and extend the treatment and management of patients with maple syrup urine disease and provide a method for study of other mutant human enzymes located in the mitochondrial membrane.

A role for thiamine in the regulation of fatty acid and cholesterol biosynthesis in cultured cells of neural origin.

Abstract: — Cultured glial (C-6) and neuronal (neuroblastoma) cells were utilized to define the role of thiamine in the regulation of fatty acid and cholesterol biosynthesis. Glial cells subjected to thiamine deficiency exhibited rates of fatty acid synthesis that were only 13% of the rates in thiamine-supple-mented cells. The decrease in fatty acid synthetic rate was accompanied by a comparable decrease in the activities of fatty acid synthetase and acetyl-CoA carboxylase, the two critical enzymes in the pathway. Immunochemical techniques demonstrated that the decrease in activity of fatty acid synthetase reflected a decrease in enzyme content and that this change in content was caused by a decrease in enzyme synthesis. The disturbance of fatty acid synthesis was exquisitely sensitive to thiamine–i.e. marked improvement was evident within hours of replenishment with only 0.01 μ/ml of thiamine. Total recovery occurred in 1–2 days. Thiamine-deficient glia also exhibited reduced rates of cholesterol biosynthesis, i.e. 60% of the rates in thiamine-supplemented cells. This effect was accompanied by a comparable reduction in activity of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, the rate-limiting step in cholesterol biosynthesis. Unlike the glial cells, the neuronal cells exhibited either no or only a slight reduction in lipid synthesis under similar conditions of thiamine deficiency. The data have important implications for the genesis of the neuropathology in states of altered thiamine homeostasis and for the mechanisms of regulation of lipid synthesis.

Red Blood Cell Transketolase Activity and the Effect of Thiamine Supplementation in Patients with Chronic Liver Disease

Abstract: Biochemical evidence of thiamine deficiency was found in 58% of patients with chronic liver disease, the incidence being higher in alcoholic than in non-alcoholic patients. Daily supplementation with high doses of thiamine hydrochloride (200 mg/day) for one week restored levels of thiamine pyrophosphate (TPP), the active co-enzyme form of thiamine, to normal in all cases. Such supplementation also stimulated synthesis of the TPP dependent enzyme transketolase. Because of the essential role of TPP as a co-factor in intermediary metabolism, it is concluded that high doses of thiamine should be included in the routine nutritional management of patients with severe chronic liver disease.

Effects of thiamine in a patient with a variant form of branched‐chain ketoaciduria

Abstract: A boy with the intermediate variant of branched-chain ketoaciduria was studied. Treatment with an amino acid mixture was discontinued at the age of 7.5 years. Reintroduction of normal protein-containing foods precipitated the biochemical abnormalities, characteristic of MSUD, which were relieved by 10 mg thiamine/day. Adaptation to this regiment developed and thiamine intake was increased to 100 mg/day, later to 1 000 mg/day. The patient developed well and had no attacks of ketoacidosis. 1-14C-leucine degradation in intact fibroblasts was 15% of controls and did not increase upon addition of thiamine to the incubation medium.

Effect of lead poisoning on the thiamine status and function in liver and blood of rats.

Abstract: Three groups of Sprague-Dawley rats were fed a thiamine deficient diet, which was supplemented by daily subcutaneous injections of a minimum requirement of thiamine, and treated with lead(II) acetate in different molar ratios to thiamine (1:1, 2:1, 10:1) for 5 and 9 months, respectively. The prolonged administration of lead(II) acetate decreases the thiamine level in lead-treated rats and diminishes the enzymatic activity of pyruvate dehydrogenase as well as that of transketolase. The thiamine level in the liver decreased by 30 to 40% compared with a reference group and the activity of the erythrocyte transketolase diminished by 5 to 40%. The level of the blood pyruvate increased by about 20% and the rate of the oxidative decarboxylation of pyruvate by liver mitochondria decreased.

Possible role of intestinal alkaline phosphatase activity in thiamine transport.

Abstract: Thiamine deficiency caused a marked decrease of intestinal alkaline phosphatase (al-Pase) activity, but had no effect on the Ca++-ATPase activity and Ca++-absorption in rats. The al-Pase activity was significantly decreased 1 h after oral administration of ethanol at 0.5 and 2.5 g/kg. In contrast, Mg++-, Ca++-and (Na+ + K+)-ATPase activities did not change after the administration of ethanol. These findings show that the al-Pase activity, unlike the Ca++-ATPase activity, is not related to Ca++-absorption. A possible role of al-Pase activity in the active transport of thiamine in the intestine was discussed.

Vitamin assay by microbial and protozoan organisms: response to vitamin concentration, incubation time and assay vessel size

Abstract: Standard vitamin calibration curves for the microbiological determination of thiamine, riboflavin, vitamin B6, vitamin B1 2, biotin, niacin and pantothenic acid were prepared to determine the minimal and maximal vitamin concentrations that may be assayed by various microorganisms and protozoa, and to determine the incubation periods required for the growth response of the microbial vitamin assays to stabilize. The effect of assay vessel size on growth of Tetrahymena pyriformis was also determined. Vitamin assays using Tetrahymena pyrijkmis were shown to have wider concentration limits than those of bacteria and yeast. Analyses based on Ochromonas danica and Ochromonas malhamensis possessed approximately equal vitamin concentration limits when compared to corresponding bacterial assay methods. Accepted vitamin assay methodology using microorganisms terminate vitamin assays when the growth response has stabilized. This study indicated that some test organisms such as Lactobacillus viridescens, (thiamine), Lactobacillus casei (riboflavin) and Saccharomyces uvarum (vitamin B6) do not achieve a point of stable response, thus requiring a defined incubation period. Tetrahymena pyriformis growth was depressed by use of small diameter assay vessels. Optimum response of this organism requires the use of flasks in which assay media, when dispensed, will have a large surface to volume ratio.

[Muricide induced by thiamine deficiency in the rats (author's transl)]

Abstract: The emotional behavioral aspects and the interaction between the changes of the polyamine contents and muricide response in thiamine deficient rats were investigated. In the thiamine deficient group, there was evidence of muricide and such increased progressively with advanced thiamine deficient feeding. This muricide was characteristic in the following respects; 1) the killer-rats did not eat but only kill a mouse and it was quite difficult to remove a sacrificed animal from the cage. 2) they bit at random into any body region of the mouse. 3) the killer-rats did not bite inanimate objects such as nails nor chalk. 4) the muricide induced by thiamine deficiency could not be suppressed by a single injection of thiamine HCl. On the 30th day of the experimental feeding, both spermidine and spermine levels in the brain of the thiamine deficient group decreased significantly as compared to the control and the pair-fed groups. Both spermidine and spermine levels were reversed to the control levels with a intraperitoneal administration of thiamine. There were no significant differences in spermine and spermidine levels between the killer-rats and non-killer-rats in the thiamine deficient group.

Studies on ATP: thiamine diphosphate phosphotransferase activity in rat brain.

Abstract: The experiments described in this paper serve as a contribution to the solution of the discrepancies which exist in the assay of ATP:thiamine diphosphate phosphotransferase activity (EC 2.7.4.15), presently in use as a tool for the diagnosis of Leigh's disease (SNE, subacute necrotizing encephalomyelopathy). The results obtained with this phosphotransferase assay can, in part, be explained by the presence of thiamine triphosphate (ThTP) in the preparation of thiamine diphosphate (ThDP) used as a substrate, by the inhibition by ATP of the ThTP phosphohydrolase activity, present in fractions of rat brain homogenates, and by the stimulation by ThDP of the ATPase activity. When [2(-14)C-thiazole]thiamine was used for the synthesis of [14C]ThTP in fractions of rat brain, it was found that after chromatographic separation of thiamine and its phosphates, 14C radioactivity could be demonstrated in the ThTP fractions, even in the absence of an enzyme source. Probably a complex is formed between [14C]thiamine and a phosphate ester which behaves chromatographically as ThTP. It is concluded that the assay system for the measurement of ThTP synthesis in its present form is, in our hands, not suitable for diagnostic purposes.

Lipogenesis in the brain of thiamine deficient rat pups

Abstract: (1) The effect of thiamine deficiency during pregnancy and lactation on lipogenesis in the brain of rat pups was determined. (2) Acetate incorporation to brain lipids in thiamine-deficient rats in vivo was no less than in pair fed control rats, apart from slightly reduced fatty acid synthesis in the cerebrum. (3) Glucose incorporation to brain lipids in vivo was considerably reduced in thiamine-deficient pups. (4) The inducible NADP dependent malic enzyme activity was increased in thiamine-deficient pup brains. (5) The synthesis of acetyl-CoA appears to be the rate limiting step in lipogenesis in thiamine-deficient pup brains.

Effect of intravenous thiamine on pralidoxime kinetics.

Abstract: Subjects were given pralidoxime chloride (5 mg/kg, intravenously) alone and again while they were receiving an infusion of thiamine hydrochloride. After the addition of thiamine: (1) overall, the urinary excretion of oxime was the same but the amount excreted in the first three hours was smaller; (2) the plasma half-life of oxime lengthened; (3) the plasma concentrations of oxime rose; and (4) the intercompartmental clearances and rate constant for elimination for oxime fell. These changes suggest that thiamine and oxime compete for a common renal secretory mechanism or that thiamine alters the membrane transport of oxime.

Wernicke's encephalopathy in infancy. Development during parenteral nutrition.

Abstract: Wernicke's encephalopathy documented by characteristic involvement of the mammillary bodies developed in a premature infant during parenteral nutrition. This case illustrates the need for thiamine supplementation in seriously ill patients with inadequate intake of the vitamin.

[Growth kinetics of thiamine requiring Candida lipolytica during thiamine limitation: the existence of a linear growth phase].

Abstract: The growth kinetics of batch cultures of the thiamine requiring Candida lipolytica 695 was investigated. It was established that a phase of logarithmic growth is followed by a linear growth phase. The time length of the linear phase and the relative increasing of the biomass during the linear phase are nearly constant and independend of the concentration of thiamine. The existence of the linear phase is discussed in connection with a constant activity of one of the two 2-ketoacid dehydrogenases as a bottle neck enzyme which needs thiamine pyrophosphate as coenzyme. Critical intracellular thiamine concentrations were calculated necessary for transition from the logarithmic to the linear phase and from the logarithmic to the stationary growth phase. Without the existence of the second critical thiamine concentration the linear growth would continue infinitely.

Water-soluble vitamins in severe liver disease.

Abstract: Biochemical deficiency of thiamine, vitamin B6, ascorbic acid or nicotinic acid occurred in 71% and 88% of patients with fulminant hepatic failure (FHF) and decompensated chronic liver disease (DCLD) respectively. Transient high plasma vitamin B6 concentrations in FHF were followed by low levels later in the illness. Although patients with DCLD of alcoholic aetiology tended to have lower circulating levels of vitamins than those with non-alcoholic DCLD, the prevalence of abnormally low concentrations did not differ. Decreased dietary nutrient intake and alcohol appeared to be less important determinants of biochemical vitamin deficiency than the presence of liver disease per se. Finally, urinary excretion of these vitamins or their major metabolites in patients with severe liver disease correlated poorly with circulating levels of vitamins.