Neonatal screening for biotinidase deficiency in Hungary: clinical, biochemical and molecular studies.

From 1989 to 2001, 1,336,145 newborns were screened for biotinidase deficiency in Hungary. Fifty-eight children with the disorder were identified as enzyme-deficient. We have characterized the clinical and biochemical features and mutations of 20 of these children. Eleven children had profound biotinidase deficiency, 7 had partial biotinidase deficiency, and 2 were found to be heterozygous for profound deficiency by mutation analysis. Seventeen different mutations were identified in this population including seven novel mutations. Six of these new mutations are missense, 245C>A, 334G>A, 652G>C, 832C>G, 1253G>C, 1511T>A, and one is a unique allelic double mutation [212T>C;236G>A]. Of five Romanian Gypsies, four were homozygous for the 1595C>T mutation and one was heterozygous for this mutation. Most of the children with profound deficiency have been asymptomatic on therapy; however, four exhibited minimal brain abnormalities, motor delay and abnormal blood chemistries. Compliance with therapy must be questioned in these cases. Of clinical importance, all of the children with partial deficiency exhibited mild symptoms at the time of diagnosis, at several weeks to months of age. These symptoms resolved following biotin therapy. This is in contrast to the experience in the United States, where the children with partial deficiency have been asymptomatic at the time of diagnosis. This finding further indicates that children with partial deficiency should be treated. The incidence of biotinidase deficiency in Hungary is more than twice that observed in a worldwide survey. These results indicate that newborn screening in Hungary is effective and warranted.

The effect of erythropoietin on platelet function in uraemic children on haemodialysis.

The effect of 1-year erythropoietin (rHu-EPO) treatment on the bleeding time, platelet aggregation, ATP and thromboxane B2 (TXB2) release, cyclic AMP (cAMP) concentration and platelet surface positive charge were studied in 8 haemodialysed children with chronic uraemia and 8 controls. The pre-dialysis haematocrit (Hct) was 0.21 + 0.01 before and 0.36 + 0.01 following 1 year of rHu-EPO therapy. At the end of this period the pre-dialysis bleeding time became normal (P < 0.05); this was associated with a significant increase in platelet aggregability (P < 0.05), ATP release (P < 0.05) and TXB2 production (P < 0.01), and with a significant decrease in platelet cAMP concentration (P < 0.01). A further increase in platelet aggregation, ATP release and TXB2 production and a decrease in platelet cAMP concentration was observed following bicarbonate haemodialysis (BHD) (P < 0.01). There was a significant positive correlation between platelet aggregation and ATP release (r = 0.78, P < 0.05), as well as platelet aggregation and TXB2 production (r = 0.68, P < 0.05). A significant negative correlation was found between platelet aggregability and cAMP concentration (r = -0.7, P < 0.05). The platelet surface positive charge, which was significantly lower in the patients than in the controls (P < 0.01), did not change during rHu-EPO therapy, nevertheless BHD resulted in a significant increase (P < 0.05), suggesting the surface charge may influence platelet aggregation. In an in vitro and an in vivo study, rHu-EPO and the higher Hct did not increase platelet aggregation directly. Long-term administration of rHu-EPO stimulated complex functional and biochemical changes in the platelets of uraemic patients, which resulted in an improved aggregability.

Laser aggregometer studies, ATP release and thromboxane B2 release and cAMP concentration of the platelets in nephrotic syndrome.

Platelet function was studied in 56 children with nephrotic syndrome, 33 were on oral prednisolone (P) treatment (group 1), while 23 were in early (< 6 months) remission (group 2): 12 on P (group 2a) and 11 not on P (group 2b), and there were 18 controls (group 3). The following tests were used: platelet aggregation with collagen in a laser rheoaggregometer; adenosine triphosphate (ATP) release: during aggregation with luciferin-luciferase in a lumiaggregometer; thromboxane B2 (TXB2) release: by radioimmunoassay; platelet cAMP concentration: by binding assay. The changes in plasma cholesterol (C) and triglycerides (TG) were compared with the platelet aggregation results. Patients in group 1 and 2 exhibited significantly higher aggregability, TXB2 release and ATP release in response to collagen than those in group 3 (p < 0.01), but there was no difference between groups 1 and 2 or groups 2a and 2b. Some differences were observed between the histological groups. Patients with IgA and SLE nephropathy displayed higher aggregability than those with minimal change nephrotic syndrome in remission (p < 0.05). The highest level was in membranous nephropathy. The platelet cyclic adenosine monophosphate (cAMP) concentration was significantly lower in groups 1 and 2 than in group 3 (p < 0.001). No differences were observed between groups 1 and 2 or between groups 2a and 2b. Plasma C and TG levels did not show any correlation with the platelet aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)

[Biochemical, clinical and genetic analysis of various aminoacidopathies (non-ketotic hyperglycemia, maple syrup urine disease, histidinemia, tyrosinemia)]. / Néhány aminoacidopathia (non-ketotikus hyperglycinaemia, jávorfaszörp-betegség, histidinaemia, tyrosinaemia) biokémiai, klinikai és genetikai elemzése.

The genetical types were classified according to the clinical findings and biochemical results in cases of 13 newborn/children suffering from various aminoacidopathies. The genetical types were: 3 neonatal and 4 infantile types were found out of 7 non-ketotic disease (MSUD) patient was infantile type with 9.1 per cent keto acid decarboxylase activity in leukocyte homogenate. Among the 3 histidinemic patients 1 was severe neonatal type and 2 cases were chronic types. The 2 treated tyrosinemic children proved to be type III. (chronic with rickets).

The effects of bicarbonate and acetate haemodialysis on platelet cyclic AMP concentration, thromboxane B2 release and aggregation.

The effects of chronic uraemia and serial acetate (HDA) or bicarbonate (HDB) haemodialysis on the aggregation, thromboxane B2 (TXB2) release and cyclic AMP (cAMP) concentration of platelets from arterial blood were studied in 14 uraemic patients (6 dialysed and 8 conservatively treated) and 10 controls. Platelets from uraemic patients, either dialysed or treated conservatively, exhibited a significantly higher cAMP level (P less than 0.005), a lower TXB2 level (P less than 0.01), and a lower aggregability (P less than 0.001) than the controls. The platelet cAMP level was more markedly decreased after HDB than after HDA (P less than 0.05). Greater increases in platelet aggregation (P less than 0.05) and TXB2 formation were observed after HDB than after HDA. The concentration of platelet cAMP and aggregability, and also the platelet cAMP and the TXB2 level showed a significantly negative correlation (r = -0.7, P less than 0.05 and r = -0.60, P less than 0.05, respectively). There was a positive correlation between the platelet-derived TXB2 and the aggregability (r = 0.67, P less than 0.05). Although most patients had secondary hyperparathyroidism, the serum parathyroid hormone level did not correlate closely with the cAMP, TXB2 or aggregation results. The dysfunction of uraemic platelets accompanied by a reduced TXB2 release may be explained by an increased cAMP and a decreased arachidonic acid availability. HDB improves the platelet function to a greater degree than does HDA.

Biochemical and ultrastructural diagnostic problems in mucolipidoses.

Biochemical and ultrastructural investigations were made in 2 children suffering from mucolipidosis type III. Among the lysosomal hydrolases the activity of beta-galactosidase and alfa-fucosidase diminished in the homogenate of the peripheral leukocytes in case I. The activity of serum and leukocyte arylsulfatase was normal. By electron microscopy typical storage organellums for mucolipidosis were detected in different biopsy materials--liver, skin, conjunctival ones--and in the cytoplasm of the peripheral lymphocytes and leukocytes. Definitive diagnosis was given by the specific electron microscopic investigations detecting the typical storage patterns for mucolipidosis.

Neonatal screening for biotinidase deficiency in east-Hungary.

There are two types of multiple carboxylase deficiency, the neonatal form with holocarboxylase synthetase defect and the late-onset form with biotinidase deficiency. We report our preliminary experiences in screening for biotinidase deficiency. In total 43,493 infants were screened for the deficiency of the enzyme biotinidase; 0.14% false positive results that necessitated requests for second blood samples and two newborns with a biotinidase defect were identified during our pilot study. The definitive diagnosis required the demonstration of enzyme deficiency in serum. Both of the patients have residual biotinidase activity: 3.59% and 7.55%. These two newborns with biotinidase deficiency are treated with daily supplementation of free biotin. According to our preliminary results biotinidase deficiency satisfies all the criteria for incorporation into the national newborn mass screening.

[Hypoxic myoglobinemia in severe neonatal disorders of adaptation to extrauterine life]. / Hypoxiás myoglobinaemia vizsgálata az újszülöttek extrauterin élethez való súlyos adaptációs zavaraiban.

In two series of newborns needing intensive care the presence, the degree and the prognostic value of myoglobinaemia was examined. In series I. of hypoxic newborns the myoglobinaemia was present even in infants requiring less than 60% O2 therapy. The serum myoglobin value was significantly higher in cases needing oxygen therapy over 60% oxygen. This was most pronounced in the critically severe and progressive cases. In series II. of 34 consecutive cases of hypoxic newborns exceeding 7 nM/l proved to have a prognostic value indicating critically severe course or fatal outcome of the disease. The myoglobinaemia observed in the present study may explain the effectivity of the peritoneal dialysis therapy introduced previously by us in severe hypoxic newborns. This possibility was supported by further observations on the transperitoneal passage of myoglobin in 4 renal hilus ligated and peritoneally dialyzed newborn piglets. In conclusion, early detection of the elevated myoglobinaemia in severely hypoxic newborns has a definite prognostic value and its degree can be used in the indication of peritoneal dialysis.

[Mass screening of neonates for histidinemia]. / A histidinaemia újszülöttkori tömegszürövizsgálata.

The authors report on newborn mass-screening for histidinemia by Guthrie bacterium inhibition assay. 51,618 newborns were screened and 805 cases with more than 5 mg% histidine blood level were found. A second test, urocanic acid thin-layer chromatography was done from these samples. According to these tests no histidinemic patient was detected. It is worth mentioning that blood histidine level exceeded 20 mg% in 55 cases from 805 newborns. Considering our screening results, the low incidence and the relatively good prognosis of the disease, a mass-screening in Hungary is not necessary.

Plasma factors influencing PGI2-like activity in patients with IgA nephropathy and Henoch-Schönlein purpura.

Plasma factors influencing vascular PGI2-like activity (PSA) were studied in 45 patients with IgA nephropathy, 18 with Henoch-Schönlein purpura, including 8 children with nephrotic syndrome, and 41 controls. The results were compared with the levels of plasma high-density lipoprotein (HDL), low-density lipoprotein (LDL) and fatty acid components of plasma phospholipids. The plasma of 38 of 45 patients with IgA nephropathy and 14 with Henoch-Schönlein purpura showed a diminished ability or no ability to support PSA. Twenty-three patients with IgA nephropathy and 10 with Henoch-Schönlein purpura exhibited an inhibitory activity against PGI2 production. The plasma HDL level was lower, while the LDL level and the LDL/HDL ratio were significantly higher in IgA nephropathy and Henoch-Schönlein purpura cases than in the controls. A high LDL/HDL ratio was associated with a low plasma PSA. The levels of arachidonic acid and its precursor were not lower in the plasma of patients than in the controls. The decreased PGI2 synthesis may play an important role in the pathogenesis of IgA nephropathy and Henoch-Schönlein purpura, but it can not be explained by reduced PG precursors. LDL may have an inhibitory, and HDL a protective effect on PGI2 synthesis.

The effects of plasma from IgA nephropathy patients on vascular prostacyclin and platelet cyclic AMP production.

The effects of plasma from 10 IgA nephropathy patients and from ten controls were studied on vascular prostacyclin (PGI2) production, the cyclic AMP (cAMP) level and the aggregation of normal platelets. The ability of the plasma to support PGI2-like activity (PSA) was significantly lower in the group of patients (18.0 +/- 13.3%) than in the controls (52.6 +/- 12.9%). The concentration of 6-keto-PGF1 alpha in the supernatant of the vascular tissue was also lower following incubation with patient plasma than with control plasma (p less than 0.001). The reduced PGI2 released by the patient plasma led to a significantly lower platelet cAMP than that following the control plasma (p less than 0.01). There was a significantly positive correlation between the 6-keto-PGF1 alpha and the plasma PSA, and also between both the plasma PSA and 6-keto-PGF1 alpha concentrations and the platelet cAMP level. These findings suggest that a vascular PGI2 defect may cause a reduced cAMP production and an uninhibited aggregation of platelets, which might play a role in the pathogenesis of IgA nephropathy.

Manifestation of angiokeratoma diffusum in a girl patient with heterozygous genotype for Fabry disease.

The case of a 16 years old girl patient having angiokeratoma corporis diffusum with Fabry heterozygosity is published together with the clinical and genetical screening investigations of the family members. The problem of the detection of the heterozygotes is discussed.

Ultrastructural study of peripheral lymphocytes and polymorphonuclear leukocytes in children with lysosomal enzymopathies and hyperlipoproteinemia.

On the basis of electronmicroscopic examinations of the peripheral lymphocytes and polymorphonuclear leukocytes (PMNL) in mucopolysaccharidosis of types I and II in Gaucher and Nieman-Pick diseases, in metachromatic leukodystrophy and in hyperlipoproteinemia, the ultrastructural characteristics are described. Pathological findings with vacuoles formations were observed in Gaucher disease and in metachromatic leukodystrophy against the preliminary literature. The ultrastructural pathological changes are reported from the first ultrastructural PMNL examinations in hyperlipoproteinemias. Electronmicroscopic analysis of the leukocytes is considered to give information equivalent in value to that from liver biopsy studies, but is advantageous in view of its non-invasive nature.

Changes of serum lipids and lipoproteins during haemodialysis treatment in dialysed chronic uraemic patients.

Changes of serum lipids and lipoproteins were determined quantitatively before and after haemodialysis in chronic uraemic patients. Serum beta-lipoprotein significantly decreased due to haemodialysis, while alpha-lipoprotein and the FFA level significantly increased. Slight correlations were observed between the applied transmembrane pressure and the serum concentration of FFA, HDL-cholesterol or total cholesterol levels. Hyper-beta-lipoproteinemia was found in 70.9 per cent and Frederickson-HLP in 33.3% of the haemodialyzed chronic uraemic patients.

The role of renal prostaglandin E as a possible modulator of cyclic AMP production in nephrotic syndrome.

Urinary prostaglandin E (PGE) and cyclic AMP (cAMP) excretions were studied by radioimmunoassay in children with nephrotic syndrome and in a control population. In cases with nephrotic syndrome there was a significant elevation in urinary PGE excretion and cAMP excretion was decreased. A positive correlation was found between urinary cAMP excretion and urinary osmolality (Uosm) and the ratio urine to plasma osmolality (Uosm/Posm); and a negative correlation between urinary cAMP excretion and urine volume. A negative correlation was observed between the values of PGE excretion and urinary cAMP. These data confirmed the role of PGE as a modulator of cAMP production, which was inhibited in the nephrotic syndrome.