Metformin, valproic acid, and starvation induce seizures in a patient with partial SLC13A5 deficiency: a case of pharmaco-synergistic heterozygosity.

SLC13A5/NaCT is a sodium-coupled citrate transporter expressed in the plasma membrane of the liver, testis, and brain. In these tissues, SLC13A5 has important functions in the synthesis of fatty acids, cholesterol, and neurotransmitters. In recent years, patients homozygous for recessive mutations in SLC13A5, known as SLC13A5 deficiency [early infantile epileptic encephalopathy-25 (EIEE-25)], exhibit severe global developmental delay, early-onset intractable seizures, spasticity, and amelogenesis imperfecta affecting tooth development. Although the pathogenesis of SLC13A5 deficiency remains not clearly understood, cytoplasmic citrate deficits, decreased energy status in neurons, and citrate-zinc chelation are hypothesized to explain the neurological deficits. However, no study has examined the possibility of specific pharmacological drugs and/or lifestyle changes synergizing with heterozygosity of SLC13A5 deficiency to increase the risk of EIEE-25 clinical phenotype. Here, we report on a heterozygous SLC13A5-deficient patient who demonstrated evidence of pharmaco-synergistic heterozygosity upon administration of metformin, valproic acid, and starvation. The report illustrates the importance of careful consideration of the potential adverse effects of specific pharmacological treatments in patients with heterozygosity for disease-causing recessive mutations in SLC13A5.

Effect of added fiber on the glucose and metabolic response to a mixed meal in normal and diabetic subjects.

10 g of guar was added to a test meal given to 11 subjects, six of whom had insulin-dependent diabetes, after an overnight fast. The addition of guar to the meal produced an overall decrease in the blood glucose concentrations after the meal in both normal and diabetic subjects. The insulin secretory response in the normal subjects was less when guar was included in the meal than when it was not. In normal subjects, but not in diabetic subjects, the addition of guar also resulted in a diminished response of lactate, pyruvate, and alanine to the test meal and a blunting of the fall in serum ketones. This is likely related to the fact that the same dose of insulin was given in the diabetic subjects with both test meals. Thus, in the normal circumstances, the diminished glycemic response to the guar-containing meal resulted in diminished insulin secretion, which was not the case with the diabetic subjects.

An enzyme specific method for estimation of pyruvic acid radioactivity in biological samples.

A method for the estimation of pyruvate radioactivity in complex biological samples is presented. After perchloric acid deproteinization, the samples were divided in two aliquots. All pyruvate was removed from the first aliquot with lactate dehydrogenase and NADH; the other was left undisturbed. Then all hydrophilic components of the sample were removed through an Amberlite XAD-7 column. The samples were treated with dinitrophenylhydrazine to obtain the corresponding hydrazones, which were retained in an Amberlite XAD-7 column and then were eluted with acetone and their radioactivity counted. The difference between both aliquots corresponded to the pyruvate radioactivity in the sample. No contamination by lactate, glycerol, glucose or amino acids was observed. The mean recovery of 5 determinations of pyruvate radioactivity was 91.3% +/- 1.0.

[The relationship between pre-exertional biochemical changes metabolic features during muscular activity]. / Sviaz' mezhdu predrabochimi biokhimicheskimi izmeneniiami i osobennostiami metabolizma pri myshechnoi deiatel'nosti

Metabolic changes during intense muscular activity and the work efficiency depend on the prestart biochemical changes. Excessiveness of these changes or, particularly, their absence decrease the work capacity. Character of the prestart reaction depends on the grade of adaptation of the organism to muscular activity; favourable, moderate prestart biochemical changes are characteristic o highly trained sportsmen. Disturbed ATP balance in the working muscles was found not to be always a limiting factor for the work capacity.

Operation of a miniature redox hydrogel-based pyruvate sensor in undiluted deoxygenated calf serum.

An amperometric sensor for the detection of pyruvate in biological fluids was formed by modifying the tip of a 0.25 mm gold wire with a layer of electrically "wired" recombinant pyruvate oxidase (POP). The sensor did not require O2 for its operation. The electroactive area of the tip of the microwire was increased by electrodeposition of platinum black. The POP was adsorbed on the platinum black and then "wired" with the cross-linked, subsequently deposited poly(4-vinylpyridine), part of the pyridine functions of which were complexed with [Os(bpy)2Cl](+/2+) and part quaternized with 2-bromoethylamine. In the resulting thin layer the POP was well "wired". When the electrode was poised at 0.4 V vs Ag/ AgCl, the sensitivity at pH 6 was 0.26 A cm(-2) M(-1) and the current increased linearly with the pyruvate concentration through the 2 x 10(-6) - 6 x 10(-4) M range. Thiamine diphosphate, flavin adenine dinucleotide, and MgCl2 were not required for the assay, but stabilized the stored enzyme electrode. Placement of a dialysis membrane (MWCO 3500) on the electrode alleviated the severe interference of ascorbate. In calf serum, the detection limit was 30 microM, suggesting that the electrode might be used in the continuous monitoring of pyruvate in hypoxic organs.

Single- and coupled-enzyme nylon tube reactors for routine determination of pyruvate and lactate in serum.

We describe the use of a continuous-flow clinical analyzer with an immobilized coupled-enzyme nylon tube reactor and an immobilized single-enzyme nylon tube reactor for routine estimation of lactate and pyruvate in serum. These reactors are incorporated into the flow system of a modified continuous-flow analyzer (Technicon AutoAnalyzer). Results for serum lactate and pyruvate by this method are compared with those by corresponding methods in which the same enzymes are used in solution, either automatically (pyruvate) or manually (lactate) performed. Routine clinical laboratory determinations with use of the coupled-enzyme system lactate dehydrogenase and alanine aminotransferase, co-immobilized in the nylon tube reactor for estimation of lactate, and lactate dehydrogenase reactors for estimation of pyruvate give reliable and reproducible results with high precision at low cost.

Relationship of blood and saliva lactate and pyruvate concentrations.

Lactate and pyruvate concentrations of blood and saliva were investigated in nine healthy young men before and after maximal exercise. Lactate and pyruvate concentrations in saliva and blood were found to be unrealted. Interpretation favored salivary gland anaerobic metabolism as the origin of salivary lactate, but the large difference of 80 mg per 100 ml between blood and saliva found after 12 min of recovery may represent a threshold value and leaves unanswered the question on the origin of lactate in saliva. Multiple sampling following the recovery period for a longer time may change the present interpretation.

Blood biochemical alterations during recovery from competitive marathon running.

The occurrence of possible prolonged alterations in plasma levels of cortisol, epinephrine, norepinephrine, selected energy metabolites, and water and electrolyte balance was assessed in six highly trained male marathon runners by measuring blood biochemical constituents on the 10th day before and for the three days following the running of a marathon. Post-race changes in hemoglobin, hematocrit, and protein levels indicated that a hemodilution had occurred, possibly as a result of elevated sodium levels. Epinephrine remained elevated until the second day. Cortisol was depressed on the first and third post-race days while variable levels were observed on the second day. No prolonged alterations were noted in norepinephrine, FFA, glycerol, triglycerides, glucose, lactate, and pyruvate.

Reconstitution of the L-lactate carrier from rat and rabbit erythrocyte plasma membranes.

1. Rat and rabbit erythrocyte plasma-membrane proteins were solubilized with decanoyl-N-methylglucamide and reconstituted into liposomes. The procedure includes detergent removal by gel filtration, followed by a freeze-thaw step. 2. The rate of [1-14C]pyruvate uptake into these vesicles was inhibited by approx. 70% by alpha-cyano-4-hydroxycinnamate and p-chloromercuribenzenesulphonate. The extent of uptake at equilibrium was not affected by the presence of these inhibitors, but was dependent on the osmolarity of the suspending medium. 3. Reconstituted bovine erythrocyte membranes, which have no lactate carrier, showed a much slower time course of pyruvate uptake, with no inhibitor-sensitive component. 4. L- but not D-lactate competed for alpha-cyano-4-hydroxycinnamate-sensitive [1-14C]pyruvate uptake.