NEWBORN-SCREENING FOR BIOTINIDASE-DEFIC1ENCY:EVALUATION ADENOSINE ACCUMULATION IN HEMORRHAGIC CERE- OF A 4YEARS NATIONAL SCREENING PROGRAM AND A REPORT ON BROSPINAL FLUID. G. Van den Berghe, S. Stevens, J. 10 PATIENTS. K.Widhalm,S.Bischof,S.Scheibenreiter; Vanhove & J. Jaeken, Department of Pediatrics. University 41 Dept.of Pediatrics,Univ. of Vienna,Austria 44 Hospital Gasthuisberg. Leuven. Belgium. Biotinidase deficiency is an autosomal recessiv disease Although adenosine (Ado) may play a critical regulatory role in which there is an inability to cleave biotin from in brain, little information is available with respect to its biocytin,because of a complete or partial deficiency of biotini- metabolism cerebrospinal fluid (CSF). normal, CSF from dase.Clinically seizures,skin rash,alopezia,developmental regres- in In cell-free sion may occur.In Austria a nationwide screening for biotinidase- children of various ages, degradation of 1 FM Ado reached only 0.028 deficiency was started in Jan.l986:325672newborns have been 2 0.010 nmol/h/ml at 25 OC (mean 2 SEM for n = 9). Both intact and screened on their filter-paper blood samples,out of these in 10 hemolysed red blood cells (RBC, - 50,00O/mm3 of test volume) patients the dignosis biot.def.has been made by a enzymatic-spec- produced no or negligible Ado when incubated in the absence of trophotometric method of Knappe.Thus,the incidence in Austria is CSF. Incubation of intact RBC with CSF similarly did not result in Ado 1:32567;data from a worldwide screening report on an incidence of accumulation. However, incubation of a hemolysate with CSF resulted 1:61067.The mean biotinidase activity has been calculated to be in buildup of Ado at rates that were proportional to both amount of 0,40+0,12nmol/min/mlpl. (normal range:4,01-7.98)this is 6,1+2,46of hemolysate and of CSF. Accumulation was enhanced in the presence the mean normal activity.6patients were classified as "profound" of the adenosine deaminase inhibitor deoxycoformycin (1pM). Further and bpatients as partial-deficient.In regard to therapeutic re- studies showed that CSF contains a 5'-nucleotidase (activity : 7.4 f gimes we treated only these patients with a biot.def. less than 1.8 nmol/hlml at 25 OC, n = 11) w~ththe kinetic characteristics of an 10%of mean normal activity. (n=6/10mgbiotin/d) .The group of child- ren with partial def. is under control up to now.Recently it has ecto-5'-nucleotidase (high affinity for AMP, inhibition by adenosine 5'- been published that also children with a residual activity betw. methylene diphosphonate, no stimulation by 2,3-bisphosphoglyce- 10+30~~shouldbe treated.Due to recent knowledge clinical symptoms rate). It is concluded that Ado may accumulate in hemorrhagic CSF as can be avoided and reversed by means of early treatment(5-l0mg a result of the combination of red cell hemolysis providing AMP, the biotin/d).Therefore a neonatal screening seems to be highly justi- presence of a membrane-derived 5'-nucleotidase, and the low activity fied and early treatment of all patients is clearly indicated. of adenosine deaminase.

DEVELOP5lESTAL CHAUGES N ERYTHROPOLEm' (Ep) PLASXd I>mSOREACTIVE CATIONIC TIIYPSIX (ICT) PHARMACOlU3ETICS IN FETAL AND NEONATAL SHEEP. PATTERN IN RdT MODEL OF CYSTIC FIBROSIS (CF)- RESEMaLANCE TO HUMANS.Weizman 2vi.Pediatri.c John A. Widness. Peter Veng-Pedersen. Nishit B. Mcdi, Rokn L. Gastroenterology,Soroka Hospita1,Ben-Gurion 42 Schmidt and David H. Chesmut. The College of Medicine, Depanments of Pediatrics and Anesthesia; and The College of Pharmacy; The 45 University, Beer-Sheva, Israel. University of Iowa. Iowa City, IA 52242 U.S.A. Plasma ICT is elevated in CF infants,before exocrine pancreatic insufficiency(EP1)develop. Ep is the primary hormone responsible for erythrocyte production throughout due to unknown mechanism.Reserpinized(RT)rats show CF- development. It has been proposed that recombinant human Ep f mEp) k used for the -like secretory defects,including EF1,but plasma ICT uement of anemia in premature hman neonates. Before doing so, it is impnant to examine pharmacokineuc panmeters during development in animals. To do so we has not been studied.This study explored plasma ICT developed a sensitive and specific mEp immunoprecipitation assay using plyclonal pattern and mechanism in this model.ICT(RI.\)in pancre- antisera. A nonsompanmental approach was employed to determine phannacokinetic -atic juice(PJ)and plasma,PJ volumedprotein and pancre- parameters in 6 fetal (FET) (125-133 d: 2.8-3.4 kg). 6 newborn (NB) (10-19 d; 5.0- -as weight were determined in rats,RT for 4 or 7 days, 10.6 ke). and 5 adult (ADULT) pregnant sheep (120130 d gest) following bolus post caerulein stimulation,vs.pair-fed controls (C). lnject!on of tracer amounts of lrsl.i~E~Results (.U *SO) demonsu~ledmore r3pld Results : 4 days 7 days '~olasma cleamce (CI).shorter terminal half-11fe(tln kla), greater plasm3 J~stnbut~on RT(n=8) C(n=8) RT(n=7) 1 C(n=8) volume (Vd), and greater steady state distribution volume (~ss)in the FET and NB groups (Table). We rotein]8.2T2.4 7.671.8 2.871.3*/ 5.472.1 PJ volume(mg/min/~) 15.621.3 4.271.6 1.670.7*1 3.1-0.6 (*PLO.Olvs.C) Compared to controls:(A) 4 days RT rats show higher plasma ICT,and similar PJ volume & ICT. (B) In 7 days RT rats.plasma ICT,and PJ volume 6. ICT, all decline. Conclusions:L.This nodel resembles human elimination and lmedistribution volume. For mEp to k effective in the treaunent of CF,as to elevated plasma ICT before EPI.Z,Elevated anemia in prematu% neonates, it is likely hat large; Mpdoses per kg than those used plasma ICT,is probably not due to ductular obstruction. in mating anemic adult patients will be needed. -3.0tner mechanisms are to be studied in this model.

SERUM IMMUNOREACTIVE ERYTHROPOIETIN IN CHILDREN WITH HUYAN CHORIONIC GONADOTROPIN RELEASE BY HUMAN TERM ACUTE LEUKAEMIA AT VARIOUS STAGES OF DISEASE, AN0 THE PLACENTA IS DYNAMICALLY REGULATED BY GLUCOSE EFFECTS OF TREATMENT. Marit Hellebostad(l), Jens Mar- Ivo A.Henrichs, Winfried ~.~ossmanithl, 43 strander(l), Sophie H. Slerdahl(Z), P. Mary Cotea(3!, 46 Reiner ~enzl,Walter H.Teller Harald E. Refsum(4). 1) Dept. of Paediatrics, Ulleval Depts. Pediatrics I and l~ynecology, Hospital, Oslo, Norway, 2) Dept. of Paediatrics, Riks- University of Ulm, D-7900 Ulm, FRG hospitalet, Oslo, Norway, 3) Clinical Research Centre, Harrow, During gestation human chorionic gonadotropin (hCG) is secreted Middx., U.K., 4) Laboratory af Clinical Physiology, Ullevil Hoapi- in an initial rise to maintain corpus luteum function. In early tal, Oslo, Norway. pregnancy, also spontaneous pulsitile hCG secretion regulated by Most children with leukaemia are anaemic at the time of diagno- a GnRH-like compound of placental origin has been described. - sis and at various times during treatment. Serum erythropoietin Question: Are there metabolic influences which regulates hCG re- (EPO) was estimated by a radioimmunoasaay method in 27 children lease? - Methods: Explants of human cerm placenta (500 mg) were with acute leukaemia (~26)or lymphoma (n:l) at diagnosis (n=16), perifused in 1 ml chambers (ACUSYST, Endotron~cs) with a flow in relation to high dose methotrexate (MTX, nrll) or cytosine ara- rate of 100 ul medium 199/min. The glucose concentrations were binoside (Ara-C, n:8), and during oral maintenance therapy with changed from 5.55 to 0 mmol/l resp. 16.0 to 5.55 mmol/l in pulses MTX and 6-mercaptopurine (n-10). At diagnosis serum EPO was in- of 16 min. The experiments lasted for 5 hours. HCG was measured creased in the children with anaemia, and inversely related to every 4 min by IRIMA. - Results: 1. In hypoglycemic conditions, haemoglobin (Hb, r=-0.94, pL0.00005). After high dose MTX, in the lowered glucose concentration was followed by significant some children serum EPO increased where Hb was unchanged or in- peaks (PULSAR analysis) of hCG reaching more than 40.9'25.2 % creased. After high dose Ara-C Hb declined, and serum EPO increa- (mean + SD) of basal level (duration 6.1'1.8 min) (n=13). 2. Re- sed markedly in all cases. During oral maintenance therapy when Hb peated lowering of glucose (max. 3 times) showed corresponding was in the normal range, serum EPO was slightly increased in some hCG bursts (height: 16 - 168 mIU/ml). 3. Decreasing glucose con- children. In conclusion, children with leukaemia respond to anae- centrations in rerifusate per se were answered by elevated hCG mia with an increase in serum EPO concentration, but in relation bursts (n=3).- Conclusion: In vitro, human placenta reacts to di- to treatment with high dose MTX and Ara-C, additional mechanisms minished glucose supply with increased hCG release. This does not may influence the EPO concentration. seem to be due to a total lack of fuel.-Supp. by DFG (He110712-3)