Evidence for the complex relationship between free amino acid and sugar concentrations and acrylamide-forming potential in potato.

Free amino acids and reducing sugars participate in the Maillard reaction during high-temperature cooking and processing. This results not only in the formation of colour, aroma and flavour compounds, but also undesirable contaminants, including acrylamide, which forms when the amino acid that participates in the reaction is asparagine. In this study, tubers of 13 varieties of potato (Solanum tuberosum), which had been produced in a field trial in 2010 and sampled immediately after harvest or after storage for 6 months, were analysed to show the relationship between the concentrations of free asparagine, other free amino acids, sugars and acrylamide-forming potential. The varieties comprised five that are normally used for crisping, seven that are used for French fry production and one that is used for boiling. Acrylamide formation was measured in heated flour, and correlated with glucose and fructose concentration. In French fry varieties, which contain higher concentrations of sugars, acrylamide formation also correlated with free asparagine concentration, demonstrating the complex relationship between precursor concentration and acrylamide-forming potential in potato. Storage of the potatoes for 6 months at 9°C had a significant, variety-dependent impact on sugar and amino acid concentrations and acrylamide-forming potential.

A rationale for cystine supplementation in severe homocystinuria.

Previous studies have shown that the thiol redox, as measured by the ratio of free/bound cyst(e)ine in unaffected individuals, remains relatively constant. In severe homocystinuria (HCU) where cyst(e)ine moieties are significantly reduced, this redox is only restored when homocyst(e)ine moieties are also taken into account. This appears to stem from an increase in the free/bound homocyst(e)ine ratio with free homocystine acting as a surrogate for free cystine. We examined these ratios in 47 patients (two with a cobalamin C defect, two with methylenetetrahydrofolate reductase deficiency, 16 with pyridoxine-responsive HCU and 27 with pyridoxine-nonresponsive HCU). Comparing free/bound homocyst(e)ine ratios to the total cysteine concentration indicates a relative increase of free homocystine as total cysteine concentrations fall below 170 micromol/L. This provides a rationale and treatment algorithm for cyst(e)ine supplementation in homocystinuria.

Impairment of the nitric oxide/cyclic GMP pathway in cerebellar slices prepared from the hph-1 mouse.

In this study, the effect of tetrahydrobiopterin deficiency on the nitric oxide/cGMP pathway has been investigated in cerebellar slices derived from the hph-1 mouse. This animal displays a partial deficiency of tetrahydrobiopterin. Basal levels of cGMP were significantly reduced (-29.5%) in the hph-1 mouse cerebellum compared to controls. Following kainate stimulation (500 microM) cGMP levels increased in both control and hph-1 preparations but were again significantly lower (-29.1%) in the hph-1 mouse. Exposure of slices to the nitric oxide donors, S-nitroso-N-acetylpenicillamine and S-nitroso-glutathione, revealed no difference in cGMP accumulation between the two groups. These findings suggest that the cerebellar nitric oxide/cGMP pathway may be impaired in partial tetrahydrobiopterin deficiency states due to diminished nitric oxide formation.

An alternative way of presenting quantitative amino acid results.

An application of computer graphics is described which enables quantitative plasma amino reports to be presented pictorially in a way which enhances the quality of the information.

3-Amino-2-piperidone in the urine of patients with hyperornithinemia.

3-Amino-2-piperidone, a delta-lactam of ornithine, has been detected in the urine of a number of patients with hyperornithinemia. The distinctive yellow colour formed with a ninhydrin-cadmium reagent should help in the detection of hyperornithinemia by urinary chromatography.

Decision support techniques for the interpretation of quantitative amino acid data.

Quantitative amino acid profiles are traditionally reported as a list of individual concentrations and reference intervals. By the use of selected examples it is shown that alternative methods of presenting and assessing these results can enhance both the quantity and quality of information. A candidate decision support tool is offered as an aid to the interpretation of amino acid profiles.

Moderately low vitamin B12 does not compromise transmethylation in adults on a free diet: implications for assessment of vitamin B12 status.

Lower reference limits for vitamin B12 are often defined solely in relation to haematological criteria. This may be misleading and there is evidence indicating that biochemical anomalies should also be considered. In 50 patients we measured creatine, as the major product of B12-mediated remethylation, to see if this helps the definition of the B12 reference interval and to investigate the possible effect of low B12 on essential transmethylation. Vitamin B12 values were grouped into six fractiles covering the range 50-500 ng/L; the corresponding creatine results were assessed by analysis of variance giving F = 0.94 and a significance of 0.466. Although no correlation between B12 and creatine was found, and therefore no obvious effect on transmethylation of low B12 concentration, this must be interpreted with respect to methionine availability. Other factors indicate that biochemical perturbations should be taken into account when defining vitamin B12 reference intervals.

Homocysteine in the context of cobalamin metabolism and deficiency states.

It is becoming increasingly clear that serum vitamin B12 (cobalamin) concentration is a dubious indicator of functional B12 status and, in contrast to long-standing convention, correlates poorly with haematological indices. This, in turn, has led to poorly defined reference intervals for serum B12. Patients presenting with neurological disturbance due to B12 deficiency are at risk of not being diagnosed if total reliance is placed on serum B12 levels and haematological parameters. Plasma homocysteine remethylation is uniquely placed at the metabolic end-point of B12 metabolism such that plasma total homocysteine is proving to be a sensitive marker of functional B12 status. Studies also show that plasma homocysteine correlates better with holotranscobalamin than serum B12. It is suggested that clinicians should cease to be guided by surrogate haematological markers when more specific tests of B12 deficiency, such as holotranscobalamin and total homocysteine, exist. These tests demand greater prevalence in routine diagnostic use.

Total plasma homocysteine as part of the routine aminogram by ion-exchange chromatography.

Ion-exchange chromatography (IEC) with ninhydrin post-column derivatisation is the only technique available for the assay of total, (free plus bound), cysteine and homocysteine which also enables the routine measurement of all other commonly occurring amino acids. IEC assay of total cysteine and homocysteine typically involves incubating buffered plasma for 60 minutes at 37 degrees C with dithiothreitol (DTT), but these assay conditions significantly extend total analysis time and compromise other amino acid values, notably glutamine and glutamate. However, it is possible to carry out the DTT reduction in plasma virtually instantaneously and without additional buffering, thus preserving the integrity of other diagnostically important amino acids. Assay precision is adequate for cardiovascular risk assessment.

Glutamine-valine index - An aid in the detection of urea-cycle disorders.

By relating the increase in glutamine to the corresponding increase in valine following protein loading it has been possible to detect carriers of OCT deficiency.

Sweat sodium related to amount of sweat after sweat test in children with and without cystic fibrosis.

The sweat concentration of sodium was found to be inversely correlated with the amount of sweat obtained after a sweat test according to the method of Gibson & Cook in children without and with cystic fibrosis. Reference intervals for sweat sodium overlapped for the two groups but two-dimensional reference distributions for the amount of sweat (range 20-440 mg) correlated with its sodium content were completely separated. The establishment of similar distributions in centres carrying out sweat tests could serve to assess the performance of this investigation at local level.

Homocysteine and cysteine - albumin binding in homocystinuria: assessment of cysteine status and implications for glutathione synthesis?

Measurement of plasma total cysteine rather than free dimeric cystine gives a better indication of cysteine status in homocystinuric patients. This is the result of displacement of cysteine from albumin by homocysteine and is related to the plasma homocysteine concentration. In control subjects the free/bound cyst(e)ine ratio was independent of albumin and total cysteine concentrations. In homocystinuric (HCU) patients both free and total cyst(e)ine values differed significantly from control values (P < 0.001) but whilst free cystine considerably overlapped control values the total cysteine concentrations were almost invariably lower. The possible consequences of this on glutathione synthesis was explored by assay of plasma total glutathione but no evidence for glutathione deficiency was found. Measurement of total cysteine, rather than free cystine, provides a better indication of cysteine status in HCU.

Late onset heterozygous ornithine transcarbamylase deficiency mimicking complex partial status epilepticus.

A 57 year old woman with post-traumatic complex partial seizures was admitted because of recurrent episodes of altered mental state over the preceding 4 years, each lasting up to 5 days. There was a history of dietary protein intolerance since childhood and two of her daughters had died in the neonatal period from unexplained encephalopathies. In hospital she developed fluctuating confusion, amnesia, and sudden episodes of unresponsiveness. An EEG was consistent with complex partial status epilepticus but there was no response to benzodiazepines. Nasogastric feeding and sodium valproate were given and shortly afterwards she lapsed into a deep coma. Blood ammonia and urinary orotate were raised, and genetic testing confirmed that she was a carrier of a mutation in exon 3 of the ornithine transcarbamylase gene (C to T at position 92). Treatment with protein restriction, carnitine, and sodium phenylbutyrate led to a full recovery over a period of 3 months. To our knowledge this is the oldest age of onset yet described in a manifesting carrier. She is the fifth patient with heterozygous ornithine transcarbamylase deficiency reported to have had a severe reaction to sodium valproate. Hyperammonaemic encephalopathy should be considered in patients of any age who experience fluctuating confusion.

Plasma total homocysteine and subarachnoid haemorrhage in a co-factor replete population.

Mild hyperhomocysteinaemia is a postulated risk factor for occlusive vascular disease, including stroke. Subarachnoid haemorrhage (SAH) has an annual incidence of 10-20 per 100,000 and accounts for 5-10% of all strokes. Measurement of plasma total homocysteine (tHcy) in a cohort of vitamin B12 and folate replete patients did not reveal any association between tHcy and the aetiology of SAH.

Quantitative analysis of small intestinal mucosa in cow's milk-sensitive enteropathy.

The appearance of the small intestinal mucosa in cow's milk protein intolerance (CMPI) was studied using quantitative morphometry. The parameters under study were the numbers of eosinophil cells in the lamina propria and epithelium, villous height, crypt zone depth, villous height/crypt zone depth ratio, and total mucosal thickness. Tracings of whole sections were analysed using a suitably programmed minicomputer linked to a digitising table. Small bowel biopsy specimens from children with untreated CMPI, from children before and after clinical relapse on cow's milk challenges, and from children with resolved CMPI were compared to each other, to those from control infants, and to those from children with coeliac disease. No change of diagnostic significance could be found in the number of lamina propria eosinophil cells, but levels of intraepithelial eosinophils were significantly increased following cow's milk challenge. Quantification of mucosal dimensions confirmed the presence of a cow's milk-sensitive enteropathy and established the finding of a thin mucosa in CMPI regardless of clinical disease activity. Mucosal thickness was not different from control values following resolution of the disease. In coeliac disease mucosal thickness was significantly greater than in CMPI (apart from young children with untreated coeliac disease whose mucosa was not thicker than that of children with untreated CMPI) but not different from control values. It is suggested that in CMPI there is a limitation in the capacity of crypt cells to compensate for the loss of villous epithelium.

Primary pyruvate dehydrogenase E3 binding protein deficiency with mild hyperlactataemia and hyperalaninaemia.

A case of pyruvate dehydrogenase E3 binding protein deficiency is reported in a 24-year-old male with encephalomyopathy. Blood lactate was only minimally elevated, as was alanine.

Blood mononuclear cell coenzyme Q10 concentration and mitochondrial respiratory chain succinate cytochrome-c reductase activity in phenylketonuric patients.

Coenzyme Q10 (CoQ10) serves as an electron carrier within the mitochondrial respiratory chain (MRC), where it is integrally involved in oxidative phosphorylation and consequently ATP production. It has recently been suggested that phenylketonuria (PKU) patients may be susceptible to a CoQ10 deficiency as a consequence of their phenylalanine-restricted diet, which avoids foods rich in CoQ10 and its precursors. Furthermore, the high phenylalanine level in PKU patients not on dietary restriction may also result in impaired endogenous CoQ10 production, as previous studies have suggested an inhibitory effect of phenylalanine on HMG-CoA reductase, the rate-controlling enzyme in CoQ10 biosynthesis. We investigated the effect of both dietary restriction and elevated plasma phenylalanine concentration on blood mononuclear cell CoQ10 concentration and the activity of MRC complex II + III (succinate:cytochrome-c reductase; an enzyme that relies on endogenous CoQ10) in a PKU patient population. The concentrations of CoQ10 and MRC complex II + III activity were not found to be significantly different between the PKU patients on dietary restriction, PKU patients off dietary restriction and the control group, although plasma phenylalanine levels were markedly different. The results from this investigation suggest that dietary restriction and the elevated plasma phenylalanine levels of PKU patients do not effect mononuclear cell CoQ10 concentration and consequently the activity of complex II + III of the MRC.