Objectives-To provide preliminary evidence that the currently employed semiquantitative method of screening for glucose-6-phosphate dehydrogenase (G6PD) deficiency can only detect infants who are totally deficient for G6PD and misses all cases of partial G6PD deficiency. Setting-General population: 2150 randomly selected blood samples from the Blood Donation Department, Speliopouleion General Hospital, Athens, Greece. Neonate population: 2000 samples from neonates (50% male; 50% female) in maternity hospitals in the greater Athens area. High risk population: a total of 545 individuals from 133 families in the Athens area, the minimum acceptance criteria being the parents and any brother or sister. Method-Blood specimens from neonates or adults were collected and either spotted and dried on special filter paper (Schleicher and Schull 2992, Darmstadt, Germany) or used in tubes after being heparinised. For the quantitative evaluation of G6PD enzyme activity, the Quantase G6PD screening kit (Quantase Limited, Perth, UK) was used. Quantase G6PD controls (Quantase Limited) were used at three levels of G6PD. These controls are rated at 24, 30, and 37°C. Alternatively, we used the Sigma G6PDH controls (Sigma Chemical Company, St Louis, USA) which are rated at 30 and 37°C. The assay was performed according to the instructions included in the kit with the modification for haemoglobin normalisation. Results-General population: 36 females who were classified as having normal enzymatic activity with the semiquantitative test, were classified as partially deficient with the quantitative test. Neonate population: using the quantitative test, the percentage of G6PD deficient neonates in this population was 5.5%, compared with 3.17% reported in routine screening using the semiquantitative method. High risk population: the quantitative method detected 28 cases of total or partial G6PD deficiency in sisters of males with known total deficiency. The semiquantitative method only detected 32% (nine out of 28) of these cases.Conclusions-A considerable amount of partially G6PD deficient female neonates (heterozygotes) are undetected and classified as having normal enzymatic activity using the semiquantitative method, which uses a cut oV of 2.1 U/g haemoglobin (Hb). The use of a fully quantitative G6PD screening kit is proposed, employing the automated haemoglobin normalisation and a cut oV of 6.4 U/g Hb. Any neonate with an activity below this mark should be regarded as G6PD deficient, and all preventive measures should be taken. (J Med Screen 2000;7:46-51)

Summary:Purpose: Valproic acid (VPA) is an effective antiepileptic drug (AED), which is associated with dose-related adverse reactions such as skin rash, hair loss (alopecia), etc. Profound as well as partial biotinidase deficiency causes dermatologic manifestations similar these. Therefore, it was of interest to evaluate serum biotinidase activity in patients receiving VPA monotherapy.Methods: Seventy-five patients with seizures, mean age, 8.6 years (±1.9 years) were divided into three groups. Group A (n ‫ס‬ 25) was treated with VPA 28.7 ± 8.5 mg/kg/24 h, group B (n ‫ס‬ 25) with 41.6 ± 4.9 mg/kg/24 h, and group C with 54.5 ± 5.8 mg/kg/24 h. Their "trough" VPA serum levels were 40.9 ± 13.2, 86.25 ± 11.5, and 137 ± 14.5 g/ml, respectively. Fifty healthy children were the controls. Patients and controls underwent clinical and laboratory evaluations including liver function data, complete blood counts, NH 3 , and so on, after 45 days of VPA treatment. Biotinidase serum levels were evaluated fluorometrically.Results: Liver function data were found elevated in the groups B and C. On the contrary, biotinidase activity was significantly statistically lowered (p < 0.001) in groups B and C (1.22 ± 1.11, 0.97 ± 0.07 mmol/min/L respectively), as compared with controls (5.20 ± 0.90 mmol/min/L). Strong inverse correlations were observed between liver enzymes and VPA blood levels with the activity of the enzyme. Additionally, no inhibitory effect on biotinidase activity was found, when the enzyme was incubated in vitro with high (1.2 mM) concentrations of the drug. Skin lesions (seborrheic rash, alopecia) were improved in our patients after biotin (10 mg/day) supplementation.Conclusions: It is suggested that VPA impairs the liver mitochondrial function, resulting in a low biotinidase activity and or biotin deficiency. Biotin supplementation could restore some of the side effects of the drug.

Aim: To evaluate selenium (Se) and copper (Cu) concentrations in Greek and Albanian immigrant mothers and in the cord blood of their newborns. Subjects and methods: From 1118 Greek and 820 Albanian mothers and from the cord blood of their neonates blood was obtained for Se and Cu measurement. Se and Cu concentrations were determined in sera with graphite furnace atomic absorption spectroscopy (GFAAAS) and atomic absorption spectrometry, respectively. In all, 30 days' nutrient intakes were evaluated in both groups. Results: Animal protein, Se and Cu intakes were poor in the Albanians vs the Greeks (Po0.001). Se concentrations in the Greek mothers (68.378.5 mg/l) and in their newborns (37.0278.9 mg/l) were found higher as compared with those in Albanian mothers (37.479.9 mg/l) and in their newborns (34.379.1 mg/l) (Po0.001). Cu levels were also found higher (Po0.001) in the Greek mothers (16877353 mg/l) and in their neonates (449787 mg/l) compared with those in the Albanian mothers (9597318 mg/l) and in their newborns (229767 mg/l). Additionally, 31.5% of neonates born to Albanian women with Se concentrations less than 28 mg/l had higher Se levels (Po0.01) than their mothers. Conclusions: The low Se and Cu levels evaluated in the Albanian mothers and their newborns could be related to their poor animal protein intake which could be the consequence of their low socioeconomic status. As an effective preventive measure, accurate dietetic strategies to assess the requirements of pregnant immigrant women for trace elements may be planned in Greece.

Bone mineral status was assessed in 48 children with phenylketonuria (PKU) (20 M, 28 F, aged 2.5‐17 y). Bone density was measured in the distal third of the right forearm using single photon absorptiometry and was expressed as ±SD with respect to age‐ and gender‐matched controls. Serum calcium (Ca), magnesium (Mg), phosphorus (P), alkaline phosphatase (ALP), parathyroid hormone and 25‐hydroxyvitamin D were measured in morning samples. The ratios of urinary Ca/creatinine (UCa/ UCr), UP/UCr, UMg/UCr and hydroxyproline (OH‐Pr)/UCr were calculated in urine samples collected over a period of 3h. Patients' data were compared with those of 50 controls (22M, 28F, aged 3‐15y). The data showed severe osteopenia (below ‐2 SD) in 22/48 patients. Bone loss was more prominent in patients over 8y old. Bone density correlated significantly with age (r=– 0:56, p < 0.001) and with Phe (r=– 0:49, p < 0.007) but did not correlate with the other biochemical indices studied. Comparing PKU children with controls, significantly higher serum calcium and magnesium (p= 0:04, p< 0.001, respectively), lower ALP (p= 0:01), higher UCa/UCr ratio (p < 0.001), lower UP/UCr (p < 0.001) and lower UOH‐Pr/UCr (p < 0.001) were found. Dietary compliance was poor in patients over the age of 8y, as only 3/22 of ≤ 8y had mean serum phenylalanine >10mgdl−1, in contrast to 21/26 in the older group. It is clear from the data that osteopenia is commonly found in PKU patients from early life. The biochemical data indicate a metabolic state of low bone turnover in PKU patients. In conclusion, a better, more restricted diet may correct osteopenia.