Search

Thiamine (vitamin B-1) is an essential component of a number of metabolic pathways and thiamine deficiency results in a progressive encephalopathy in both humans and animals. Confirming thiamine deficiency is problematic and relies on demonstrating reduced red blood cells transketolase activity, or indirect methods including urinary organic acid analysis and dietary analysis. The characteristic and selective vulnerability of different brain regions in carnivores has been demonstrated by magnetic resonance (MR) imaging in the dog and cat as an aid to diagnosis. A 2-year-old, female, domestic shorthair cat was presented with an acute onset of seizures and ataxia. MR imaging was consistent with thiamine deficiency and supplementation resulted in a progressive clinical improvement. Repeated MR imaging 4 days after starting thiamine supplementation revealed near complete resolution of the MR abnormalities. Repeated MR imaging following appropriate therapy may be useful to further confirm thiamine deficiency.

The objective of this study is to investigate the effect of B vitamins-fortified foods on primary school children. A controlled trial was conducted in 101 normal primary school children aged 9-11 years. They were randomly assigned to supplemental control group (S-control, n=36), riboflavin supplementation group (+riboflavin 0.625 mg/day, n=32), and B vitamin compound supplementation group (+riboflavin 0.625 mg/day, +thiamin 0.512 mg/day, +nicotinic acid 0.365 mg/day, +folic acid 0.13 mg/day, n=33) based on school classes. Urinary riboflavin excretion and erythrocyte glutathione reductase activity coefficient (EGRAC) along with erythrocyte transketolase activity (ETKA) were used to evaluate B vitamin levels in the children. AYP index, an index reflecting the brain performance ability, was chosen to assess the children's study abilities. Health education was carried out to help children and their parents adopt scientific dietary concepts. The urinary riboflavin excretion was higher in two supplementation groups (435.24 +/- 153.3 microg/g creatinine, 374.6 +/- 144.6 microg/g creatinine) than in S-control group (235.1 +/- 86.2 microg/g creatinine). Average values of EGRAC were lower in two supplementation groups (0.90 +/- 0.11, 0.80 +/- 0.10) than in S-control group (1.08 +/- 0.25). At the same time, the percentage of thiamine pyrophosphate (TPP%) decreased from 63.69 +/- 28.04 to 42.16 +/- 16.31 in B vitamin compound supplementation group. Meanwhile, AYP index increased at the end of the supplementation in two supplementation groups. B vitamins supplementation can significantly increase B vitamin level in children. Biochemical activities of riboflavin and thiamin can improve with the intake of fortified foods. The effect of B vitamin compound supplementation is better than that of single riboflavin supplementation when the effect of riboflavin's biofunction is considered. In addition, micronutrient supplementation appears to assist children's study abilities.

Metabolic mapping of enzyme activities (enzyme histochemistry) is an important tool to understand (patho)physiological functions of enzymes. A new enzyme histochemical method has been developed to detect transketolase activity in situ in various rat tissues and its ultrastructural localization in individual cells. In situ detection of transketolase is important because this multifunctional enzyme has been related with diseases such as cancer, diabetes, Alzheimer's disease, and Wernicke-Korsakoff's syndrome. The proposed method is based on the tetrazolium salt method applied to unfixed cryostat sections in the presence of polyvinyl alcohol. The method appeared to be specific for transketolase activity when the proper control reaction is performed and showed a linear increase of the amount of final reaction product with incubation time. Transketolase activity was studied in liver, small intestine, trachea, tongue, kidney, adrenal gland, and eye. Activity was found in liver parenchyma, epithelium of small intestine, trachea, tongue, proximal tubules of kidney and cornea, and ganglion cells in medulla of adrenal gland. To demonstrate transketolase activity ultrastructurally in liver parenchymal cells, the cupper iron method was used. It was shown that transketolase activity was present in peroxisomes and at membranes of granular endoplasmic reticulum. This ultrastructural localization is similar to that of glucose-6-phosphate dehydrogenase activity, suggesting activity of the pentose phosphate pathway at these sites. It is concluded that the method developed for in situ localization of transketolase activity for light and electron microscopy is specific and allows further investigation of the role of transketolase in (proliferation of) cancer cells and other pathophysiological processes.

A method for measuring erythrocyte transketolase activity (ETKA) is described that is precise, economical of reagents and capable of high throughput with partial or full automation. The transketolase-dependent transformation of ribose-5-phosphate, yielding glyceraldehyde-3-phosphate, is linked via ‘indicator enzymes’ to glycerol production and NADH consumption, the latter being followed fluorimetrically. Conditions under which ETKA and ‘TPP Effect’ are derived have been examined and optimised. Values are comparable with those obtained by other methods but a relatively narrow adult reference range for ETKA is observed. Data are presented for optimal preparation and storage of samples.

HPLC measurement of the major physiological form of thiamin, thiamin diphosphate, in erythrocytes (ETDP) has been assessed as an indicator of thiamin status and compared with erythrocyte transketolase activity (ETKA) before and after activation with exogenous thiamin diphosphate (TDP effect). The comparison was made by following the response of the parameters to incubation of erythrocytes with thiamin and by measurements in alcoholic, elderly and student groups. All parameters were responsive to the thiamin content of their environment, but ETDP was at least as sensitive as TDP effect and more sensitive than ETKA in reflecting thiamin uptake and utilisation by erythrocytes. There was a better correlation between ETDP and ETKA than between ETDP and TDP effect. All the groups showed a significant positive correlation between ETDP and ETKA, but only the elderly and the alcoholic groups showed a significant negative correlation between ETDP and TDP effect. The lack of correlation between ETDP and TDP effect in the student group may be due to the relative imprecision of TDP effect measurements at the lower end of the reference range. ETDP is more stable than ETKA in frozen erythrocytes, is easier to standardise and is not affected by various factors which can influence the enzyme assay. Measurement of ETDP is, therefore, recommended for first line assessment of thiamin status.

The effect of a 4-week thiamine deficiency on gluconeogenesis from xylitol, lactate, pyruvate, dihydroxyacetone, and fructose in isolated rat hepatocytes was investigated. Xylitol is metabolized via transketolase, a thiamine-requiring enzyme, whereas the remaining substrates are converted to glucose via enzymes which do not require thiamine. At 7 days, transketolase activity was 81% of the value obtained with hepatocytes isolated from pair-fed control rats and gluconeogenesis from all the above substrates was normal, indicating an excess of transketolase activity present. By 14 days, however, gluconeogenesis was depressed (65-88% of control). At 28 days, both transketolase activity and gluconeogenesis from xylitol were markedly depressed (32 and 73% of control, respectively), but moreover, those substrates not utilizing thiamine-requiring enzymes for gluconeogenesis were equally or more greatly depressed in their gluconeogenic rates (42-70% of control). These data indicate (i) transketolase activi...