Met 358 to Arg mutation of alpha 1-antitrypsin associated with protein C deficiency in a patient with mild bleeding tendency.

The molecular defect responsible for a dramatic prolongation of all standard clotting tests discovered in a 15-yr-old boy has been identified. Initial investigations revealed the presence of an activated Factor X (Factor Xa) and thrombin inhibitor which copurified with alpha 1-antitrypsin (alpha 1-AT), thereby suggesting the occurrence of an alpha 1-AT variant similar to alpha 1-AT Pittsburgh. This was confirmed by dot-blot analysis and direct sequencing after amplification by the polymerase chain reaction. A G to T transition at nucleotide 10038 results in the substitution of Met to an Arg, converting alpha 1-AT into an Arg-Ser protease inhibitor (serpin) that inhibited thrombin and Factor Xa more effectively than antithrombin III. Surprisingly, there was no bleeding history in the proband. The common mutation Z, which may explain a reduced expression of the allele bearing the Arg 358 Met mutation, was not observed in the propositus' DNA. To exclude the presence of another mutation, the coding regions and intron/exon junctions were sequenced. No other mutation was found. Recently, the patient experienced his first hemorrhagic episode at the age of 17. The level of the abnormal inhibitor had increased twofold 2 mo before. The large decrease in protein C concentration may account for the mild bleeding tendency in this case, despite the presence of the alpha 1-AT Pittsburgh mutation. An abnormal protein C pattern was observed in patient's plasma, suggesting that the circulating deficiency might be due to a deleterious effect of the abnormal inhibitor on both intracellular processing and catabolism of protein C.

Catabolism of human fibrinogen fragment D in normal subjects and patients with liver cirrhosis.

The catabolism of human fragment D, (FgD), obtained by plasmin digestion of fibrinogen has been investigated in normal subjects and patients with liver cirrhosis and the results compared with those obtained for fibrinogen (Fg). Fg was labelled with I-125 and Fg D with I-131 using the chloramine T method. The plasma disappearance curves of both labelled proteins fitted a two exponential curve. In controls the plasma clearance rate of Fg D was greater than that of Fg as shown by the marked difference between the half-lives of these two tracers: 8,9 and 83,5 hours for Fg D and Fg respectively. The fractional catabolic rate of Fg D was 3.38 times the plasma pool per day. In nine patients with liver cirrhosis, catabolism of Fg was not modified. In contrast, catabolism of Fg D was significantly reduced with a half life of 13.0 hours and a low fractional catabolic rate. These results suggest the role of the liver in the catabolism of Fg D in man.

MR imaging of hemophilic arthropathy of the knee: classification and evolution of the subchondral cysts.

Magnetic resonance imaging was performed to assess the subchondral bone of the knee in 64 patients with severe hemophilia A. Using this method, subchondral cysts could be detected and evaluated. We separated the cysts into four classes (LL, LH, HH, C) depending on their signal intensities on T1- and T2-weighted images. Follow-up studies, performed on 25 patients during a period of 10-30 mo after the initial examination, suggest that these four classes of cysts correspond to four successive stages. Morphological changes within a class of cysts were also observed. Although histological confirmation was not possible, this study demonstrated that magnetic resonance imaging allows a better understanding of the pathophysiology and the natural evolution of the subchondral cysts in hemophilic arthropathy.

[Effect of ranitidine on secretion of gastric intrinsic factor and absorption of vitamin B 12]. / Effet de la ranitidine sur la sécrétion de facteur intrinsèque gastrique et sur l'absorption de la vitamine B12.

The effects of ranitidine, a new potent histamine H2-receptor antagonist, on gastric intrinsic factor (IF) secretion and protein-bound cobalamin absorption were evaluated in 6 patients with duodenal ulcer, before, during and after discontinuation of ranitidine therapy. Oral ranitidine (150 mg twice a day) resulted in a non significant decrease of IF concentration and IF output but was responsible for malabsorption of protein-bound cobalamin. This malabsorption was reversible upon discontinuation of ranitidine. These results indicate that occurrence of cobalamin deficiency cannot be excluded during long-term ranitidine treatment and emphasize the need for careful follow-up in these patients.

[Erythrocyte indices and iron stores in cord blood]. / Paramètres érythrocytaires et réserves en fer dans le sang du cordon.

BACKGROUND: Reevaluation of normal red cell values in the cord blood is necessary since these values were established before the use of automated analysers and the introduction of iron supplementation during pregnancy. POPULATION AND METHODS: Cord blood samples from all healthy babies from Monday to Friday for 2 months have been analysed for red cell count and red cell indices, hemoglobin concentration, hemoglobin electrophoresis, serum and erythrocyte ferritin concentration. The results were compared for the mothers to age, parity and duration of iron supplementation and for the neonates to gestational age and birth weight. RESULTS: One hundred and fifty-nine neonates were studied; 87% of their 154 mothers had been given iron during pregnancy. Mean red cell indices were found to be slightly below those of previous reports in the literature. They were associated with mean ferritin concentrations of 135 micrograms/l and 348 ag/cell for plasma and erythrocytic ferritin, respectively. CONCLUSION: There were no correlations between red cell indices and ferritin values in such an iron-repleted population.

The Rose Bengal test in neonatal cholestasis: diagnostic and prognostic value.

131I Rose Bengal(131IRB) studies were performed in 73 infants with extrahepatic biliary atresia (EHBA) and in 37 with intrahepatic cholestasis of various origins. Fecal 131IRB excretion of less than 10% ("complete' cholestasis) was observed in EHBA but also in some patients with either paucity of intrahepatic bile ducts (syndromatic type) or with alpha-1-antitrypsin deficiency. Seventy one 131IRB tests were also performed 3 to 8 weeks postoperatively in children operated on for EHBA. Fecal 131IRB excretion more than 15% was present in 27 out of 34 cases who were later completely jaundice free and in only one out of 37 cases where no bile flow restoration occurred. These results indicate that complete cholestasis in infants can be observed in some types of intrahepatic cholestasis, as well as in EHBA, and show that a post-operative 131IRB test is a reliable means of predicting complete restoration of bile flow in EHBA.

Erythrocyte ferritin in human neonates: maternofetal iron kinetics revisited.

Serum and erythrocyte ferritin concentrations were measured in 369 healthy term neonates and their mothers, with a view to understanding maternofetal iron kinetics. Erythrocyte ferritin concentrations in the neonates were on average 100 times higher than maternal values, while those of serum ferritin were about 10 times higher. Maternal and fetal iron status were correlated in the overall study population. Serum and erythrocyte ferritin values were also correlated with each other in both the mothers and neonates. To study the influence of iron status on these conditions, we formed three groups of mother-child pairs of equal size, based on maternal iron status (serum ferritin). The correlations found in the overall population were not always present in each subgroup. In the infants born to mothers with adequate tissue iron reserves (serum ferritin > 12 micrograms/I), a statistical link was found between serum and erythrocyte ferritin levels. When the mothers had optimal iron status (serum ferritin > or = 22 micrograms/I), there was a correlation between the neonatal and maternal erythrocyte ferritin levels. No such links were found when the mothers had low tissue iron stores (serum ferritin < 12 mg/I). These data point to special iron kinetics during fetal life and the influence of maternal iron status.

Role of bilirubin overproduction in revealing Gilbert's syndrome: is dyserythropoiesis an important factor?

Gilbert's syndrome was diagnosed in 37 patients with unconjugated hyperbilirubinaemia without overt haemolysis or structural liver abnormality, who had a marked reduction in hepatic bilirubin UDP-glucuronosyltransferase activity (B-GTA) (as compared with that of 23 normal subjects). No significant correlation existed in these patients between serum bilirubin level and the values of B-GTA, thus suggesting that factors other than a low B-GTA must influence the degree of hyperbilirubinaemia in Gilbert's syndrome. Studies of 51Cr erythrocyte survival and 59Fe kinetics in 10 unselected patients demonstrated slight haemolysis in eight, whereas mild ineffective erythropoiesis was suggested in all from a low 24-hour incorporation of radioactive iron into circulating red cells. This overproduction of bilirubin resulting from mild haemolysis and perhaps dyserythropoiesis might reflect only an extreme degree of the normal situation. It certainly contributes to the hyperbilirubinaemia of Gilbert's syndrome and may play a major role in the manifestation of this condition.

HBV infection in pediatric liver transplantation.

When hepatitis B virus (HBV) infection precedes orthotopic liver transplantation (OLT), an important issue is to avoid post-OLT HBV infection of the transplanted liver. When HBV infection is found after OLT, the main objective is to prevent the complication of cirrhosis. A study of HBV infection in 162 liver-transplanted children followed at Hôpital de Bicêtre is presented here. One hundred forty-one of these children were completely vaccinated against HBV; 89% of them still disposed of protective titers of serum antibodies to hepatitis B surface antigen (anti-HBs) (> 10 IU/L) 6-60 months after OLT. Four children had a HBV-associated liver disease that required OLT. In these children a passive immunoprophylaxis maintaining serum anti-HBs levels > 100 IU/L allowed three of them to remain free of serum hepatitis B surface antigen (HBsAg) and HBV-DNA during follow-up at 10, 20, and 36 months. Four other children have been found to be HBsAg+ after OLT, but it was not clear whether the source of HBV was a reactivation or a de novo infection. Low doses of cyclosporin and prednisone were administered, in order to avoid the complications of HBV infection. Liver needle biopsies in three of these patients 37, 42, and 46 months after OLT showed a moderate chronic active hepatitis and mild fibrosis. Immunostaining for HBs and hepatitis B core (HBc) antigens indicated active viral replication.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of folate deficiency on vitamin B12 absorption.

In three different groups of patients presenting severe folate deficiency (purely nutritional folate deficiency, chronic alcoholism or various intestinal diseases), vitamin B12 absorption has been tested by Schilling test in order to investigate the possible effect of folate deficiency on B12 absorption. Following this study, it appears that folate deficiency in itself, even severe, cannot induce vitamin B12 malabsorption, since in the first group, consisting of malnourished old people, the Schilling test was always normal even in those who presented a lowered B12 serum level; in the group of patients with intestinal disease, the Schilling test was abnormal as expected in some patients; 3 alcoholics out of 12 presented a malabsorption of B12 on the Schilling test; the follow-up of 2 of them exhibited complete correction after normal diet and alcohol suppression. Consequently, folate deficiency does not seem responsible for a secondary B12 malabsorption unless another etiological factor is present, such as alcohol or ileopathy.

[Iron deficiency in infants. Study of risk factors]. / Carence en fer chez le nourrisson. Etude des facteurs de risque.

A study of iron deficiency was carried out in two districts in Paris among 207 children aged 10 months recruited from two well-baby out-patient clinics. The main results are as follows: 12% of children were anemic, ferritin was low in 21% and 40% had a borderline deficiency, as shown by a decrease in siderophilin saturation. The average daily milk intake was greater among the immigrants, but this group, being mainly fed whole cow milk with no iron supplementation, had a lower iron intake overall. A positive correlation was found between the percentage of ingested iron-supplemented formulas and the mean corpuscular volume or serum iron levels. Iron deficiency was more frequent in children born to multiparous mothers and in the absence of any iron supplement during pregnancy. Weight gain was inversely correlated to ferritin levels, suggesting a major role of fast growth on iron metabolism.

In vitro effect of duodenal juice on R binders cobalamin complexes in subjects with pancreatic insufficiency: correlation with cobalamin absorption.

Absorption of cobalamin free or bound to chicken serum was assessed in nine patients with pancreatic insufficiency. Simultaneously the in vitro effect of duodenal juice collected from six patients and seven controls was tested on labelled cobalamin complexed to chicken serum or to R salivary binder. Malabsorption of free cobalamin was observed in one of nine patients and in four of nine patients when cobalamin was administered bound to chicken serum. The in vitro effect of duodenal juice on cobalamin complexed to chicken serum or to R salivary binder was studied: the percentage of free cobalamin released was significantly decreased in pancreatic insufficiency compared with controls whatever the binder used; the degradation of R salivary binder was different in pancreatic insufficiency and in controls. Despite the in vitro abnormalities observed in pancreatic insufficiency, these did not correlate with the in vivo absorption of cobalamin which was often normal in our patients.

Detection of hepatitis B virus DNA in serum by a simple spot hybridization technique: comparison with results for other viral markers.

A simplified spot method for determination in serum of hepatitis B virus DNA (HBV DNA) by molecular hybridization is proposed. For simultaneous testing of 30 serum samples, it reduced to about 1 hr the duration of the steps preceding hybridization proper. The method also greatly reduced the loss of DNA during these steps and allowed more sensitive detection in samples of only 25 or 50 microliters. HBV DNA was determined in 181 serum samples by this method, and the results were pooled with 67 previous determinations by the Southern blot technique. Results for the pool were then compared to those obtained with radioimmunoassay for serological HBV markers. Ninety-six of the 248 samples were HBV DNA positive. Eleven others gave variable or inconclusive results, probably due to low viral particle titers. Seventy-two HBsAg- and HBeAg-positive sera contained HBV DNA, confirming that HBeAg is a marker of active viral replication. Fourteen other HBsAg- and HBeAg-positive sera, obtained from eight patients, were either HBV DNA negative or oscillated between negative and positive, or, again, were weakly positive; serological follow-up in 7 patients showed seroconversion to anti-HBe in 5, 3 of which became HBsAg negative. Eight of the HBsAg-positive sera were negative or borderline for HBeAg but contained HBV DNA and may, therefore, have been infective; seven of these sera had anti-HBe. Six HBsAg-negative sera contained HBV DNA and may also have been infective; five of these exhibited HBV antibodies. These results indicate that molecular hybridization not only provides a more sensitive and direct method for detecting hepatitis B virus in serum but also defines additional serological patterns with predictive or epidemiological value.

Neonatal thrombocytopenia and hidden maternal autoimmunity.

Severe transient thrombocytopenia was observed in 17 newborns delivered by 11 mothers who persistently exhibited normal platelet counts and had no previous immunological or platelet disorder. Maternofetal alloimmunization as well as other causes for fetal thrombocytopenia were ruled out. A specific circulating autoantibody directed against the platelet glycoprotein Ib/IX complex was found in 10/11 mothers and in 3/4 offspring. After delivery, maternal compensated thrombocytolysis and/or hypersplenism were demonstrated by 111Indium labelled platelet studies in 10/11 women. The association of a compensated thrombocytolysis and of a specific maternal circulating antiplatelet autoantibody makes likely the diagnosis of mild maternal chronic autoimmune thrombocytopenic purpura (AITP) only expressed by the fetal or neonatal thrombocytopenia. The finding of anti-IbIX autoantibody in control women who gave birth to neonates without thrombocytopenia raises the question of the incidence and clinical significance of such antibodies which could be natural autoantibodies as well as associated with compensated thrombocytolysis or with overt immune thrombocytopenia. The cause of the discrepancy between the fetal and maternal platelet status, which could be explained by a different conformational state of fetal as compared to adult GP Ib/IX, deserves further investigation.

[Role of iron deficiency in anemia in pregnant women in Mali]. / Place de la carence martiale dans l'anémie de la femme enceinte au Mali.

The epidemiological characteristics of iron deficiency anemia after radioimmunoassay of serum and erythrocyte ferritin were evaluated in 209 Malian women at the time of delivery in a maternity unit in Bamako, Mali. The incidence of iron deficiency anemia was high (36.8%). This incidence did not reflect socio-economic status, nor even any particular obstetric history, but was significantly higher in younger mothers (aged under 26). The severity of anemia was such that 2.4% of women would require a blood transfusion post-partum. It looks therefore desirable, in Mali, to screen routinely and to ensure the prevention of iron deficiency in adolescent girls and to include, in the prevention of anemia of pregnant women, routine iron supplements from the beginning of pregnancy.

Pregnancy-associated thrombocytopenia revisited: assessment and follow-up of 50 cases.

Thrombocytopenia detected during pregnancy addresses the issue of its mechanism and of the possible occurrence of neonatal thrombocytopenia. To further investigate these issues, 50 women referred to us because of thrombocytopenia detected during pregnancy (platelet count, <150 x 10(9)/L), were extensively studied, as well as their offspring. Among these thrombocytopenic women, we used the threshold of 70 x 10(9)/L to differentiate between mild and severe thrombocytopenia. Whatever the severity of thrombocytopenia, we found biological features of an autoimmune disorder in 48% of the women, and chronic thrombocytopenia in 55%. A familial thrombocytopenia was evidenced in 1 case. These 50 women gave birth to 63 neonates, among whom 24 were thrombocytopenic, either at birth or during the first week of life. Neonatal thrombocytopenia could only be predicted in multiparous women, on the basis of previous neonatal thrombocytopenia in older siblings, and/or when maternal platelet life span study, performed before pregnancy, had evidenced an autoimmune thrombocytopenia (AITP)-like profile. These results suggest that, in case of pregnancy-associated thrombocytopenia, familial and immunological studies, combined with postdelivery iterative platelet counts, should be performed to properly characterize the thrombocytopenia. Moreover, the platelet count of the neonate should be carefully assessed at birth and during the following days, a platelet life span study should be performed after delivery in the mother, because these two parameters are likely to bring valuable information regarding the forthcoming pregnancies and the risk of neonatal thrombocytopenia.