Determination of the in vivo effects of prednisone on Bcl-2 family protein expression in childhood acute lymphoblastic leukemia.

Glucocorticoid resistance is often associated with treatment failure in children with acute lymphoblastic leukaemia (ALL) but the underlying molecular mechanisms are still unclear. In 30 consecutive children with ALL treated with prednisone we determined changes in the expression of Bcl-2, Bax and Bcl-xl proteins in leukemic lymphoblasts and related these to clinical features and rate of prednisone-induced apoptosis. The apoptotic index increased after prednisone therapy in 24 of the 30 patients. At diagnosis, we detected expression of Bcl-2 and Bcl-xl protein in 28 samples, while Bax expression protein was detected in 21 of the 30 patients. Prednisone treatment induced a decrease in Bcl-2 and Bcl-xl levels in 17 and 16 of the 28 patients, respectively, while Bax protein increased in 14 of the 21 patients. Twenty of the 30 patients studied were considered to be good prednisone responders, whereas 10 were poor responders. We observed a statistically significant decrease only for Bcl-xl protein expression in T phenotype ALL, in the poor responder group and in patients with >20000/mm(3) white cell count (WBC) at diagnosis. These data suggest a role of Bcl-xl in the mechanisms of protection of leukemic cells from apoptosis induced by glucocorticoids (GCs).

Hematopoietic stem cell transplantation in childhood: report from the bone marrow transplantation group of the Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP).

BACKGROUND AND OBJECTIVE: Transplantation of hematopoietic stem cells from different sources is being increasingly used to treat a variety of diseases in children. Transplant procedures and indications have changed considerably during recent years. Monitoring of information about these changes is useful for interpretation of nationwide collected data. DESIGN AND METHODS: Since 1985, Centers belonging to the AIEOP (Associazione Italiana Ematologia Oncologia Pediatrica), performing hematopoietic stem cell transplants (HSCT) in children, and members of the AIEOP-Bone Marrow Transplant (BMT) Group annually report data on their transplant activity to the AIEOP-BMT Registry employing specially prepared patient-oriented forms. RESULTS: From January 1985 to December 1998, a total of 2,474 bone marrow (BM), peripheral blood (PB) or umbilical cord blood (CB) transplants were reported: 1,296 (52%) were allogeneic (Allo) and 1,178 (48%) autologous (Auto) transplants. These transplants were performed in 19 Italian Centers on 2,249 patients aged less than 17 years. Among Allo-transplants, 1,198 (92%) were performed using BM progenitor cells, whereas 49 (4%) CB, 42 (3%) were PB, 4 BM plus PB, and 3 BM plus CB allografts; they were performed using HLA-identical sibling donors in 867 cases (67%) and alternative donors (i.e. partially-matched relatives or unrelated donors) in the remaining 429 (33%) cases. Allogeneic transplants were performed on 786 (67%) patients with malignancy and on 395 (33%) patients with non-malignant disorders. In the last 6 years, the number of Allo-transplants per year exceeded that of Auto-transplants. Of the Auto-transplants, 775 (66%) were performed using BM, and 403 (34%) using PB alone or combined with BM hematopoietic stem cells. Indications for Auto-BMT were myelo-lymphoproliferative disorders in 524 (49%) cases, solid tumor in 533 (50%) cases and non-malignant disease in 11 (1%) cases. In the last 5 years, the use of PB for autografts has increased from 7% to 70%. INTERPRETATION AND CONCLUSIONS: These data reflect the development and present status of HSCT in Italy and provide a basis for patient counseling and health care planning.

Interstitial 9q deletion is associated with CD7+ acute leukemia of myeloid and T lymphoid lineage.

We report the clinical, hematological and immunophenotypic characteristics from four cases of acute leukemia with interstitial deletion of chromosome 9, ie del(9)(q12-q22), as a single chromosomal abnormality. Three patients had acute myeloblastic leukemia (AML) and one T origin acute lymphoblastic leukemia (ALL). According to FAB classification, blasts were classified as M1 (two patients), M2 (one patient), and L2 (one patient). In two out of three AML cases a myelodysplastic syndrome, one AREB-t and one AREB diagnosed 6 and 11 months before respectively, preceded the onset of AML. Morphological examination showed dysgranulopoiesis, dyserythropoiesis and cytoplasmic vacuoles in two AML patients, while a strong positivity to myeloperoxidases was observed in all AML cases. As concerns immunophenotypic findings, blast cells from two of three AML patients expressed CD7 and CD34, while those from the T-ALL case displayed CD33 and CD34 along with CD7. These observations suggest that del (9q) is associated with CD7+ acute leukemia of myeloid or lymphoid lineage.

Treatment of EBV-induced lymphoproliferative disorder with epipodophyllotoxin VP16-213.

A case of haemophagocytic lymphohistiocytosis consistent with X-linked lymphoproliferative disorder is described. Remission was observed after administration of VP-213, a cytotoxic drug generally used to treat histiocytosis. The child is currently in good clinical health.

Immunologic and intestinal permeability tests as predictors of relapse during gluten challenge in childhood coeliac disease.

Fifteen children with an initial diagnosis of coeliac disease underwent gluten challenge either because they had never had a jejunal biopsy or because they had had one during the first 2 years of life. The challenge was preceded by a biopsy; clinical symptoms, the cellobiose/mannitol permeability test, and gliadin and endomysial antibody measurement were used to determine the timing of the confirmatory biopsy: it was performed if one test result was repeatedly abnormal or two results were concomitantly abnormal. Gliadin antibodies increased early (already 7 days after the reintroduction of gluten to the diet), but in many cases they returned to normal values thereafter. Increased intestinal permeability to sugars and even more positivity of endomysial antibody were good predictors of histologic relapse. The sequential use of laboratory tests during gluten challenge may significantly shorten its duration.

Iron absorption and iron deficiency in infants and children with gastrointestinal diseases.

Iron status, iron absorption, and intestinal blood loss were studied in 199 children undergoing diagnostic evaluation for suspected malabsorption. Evaluation of iron status included hematological indices, serum ferritin, and transferrin saturation. Iron absorption was assessed by the increment of serum iron after an oral iron load. Iron deficiency was common among patients affected by malabsorptive states, such as celiac disease (84%), cow's milk intolerance (76%), Crohn's disease (72%), and giardiasis (64%), whereas it was less common among patients with postinfectious enteritis (41%) and chronic nonspecific diarrhea (11%). Intestinal blood loss was seen only in patients with Crohn's disease and cow's milk intolerance, irrespective of iron nutritional status. On the other hand, iron malabsorption was very common, affecting 85-95% of the iron-deficient patients in all diagnostic groups, except in chronic nonspecific diarrhea. Iron malabsorption was less common among patients with adequate iron nutritional status than in those with iron deficiency. Iron malabsorption appears to play a major role in the pathogenesis of iron deficiency in patients with malabsorption. The iron absorption test shows greater sensitivity as a screening test for upper intestinal malabsorption than the D-xylose absorption test.

Molecular basis of chronic non-spherocytic haemolytic anaemia: a new G6PD variant (393 Arg----His) with abnormal KmG6P and marked in vivo instability.

More than 80 genetic variants of glucose-6-phosphate dehydrogenase (G6PD) are associated with chronic non-spherocytic haemolytic anaemia (CNSHA). In order to help clarify the molecular basis of this association, we have carried out a detailed biochemical and genetic characterization of two G6PD deficient brothers affected by CNSHA. The G6PD from the two patients has altered electrophoretic mobility, abnormally elevated Michaelis constant (Km) for G6P, and extreme instability in vivo and in vitro. By comparison with published information we found that this is a new G6PD variant which we have designated G6PD Portici. The entire coding region of the gene has been sequenced, and a single point mutation, a G----A transition, was found at position 1178 in exon X, causing a substitution of histidine for arginine at residue 393 in the polypeptide chain. By polymerase chain reaction (PCR) amplification followed by diagnostic restriction enzyme analysis and allele-specific oligonucleotide hybridization we have demonstrated the inheritance of this mutation in the patient's family. Our results support the notion of a causative link between this mutation in the G6PD gene and CNSHA. Our data, in combination with previous data in the literature, suggest that the three-dimensional structure of G6PD is such as to cause interaction in the binding of its two substrates, G6P and NADP.

Discriminant analysis for the diagnosis of childhood celiac disease.

A new statistical approach to the analysis of laboratory data has been introduced to optimize the use of absorption tests and gliadin antibody measurement for the diagnosis of childhood celiac disease. Serum antigliadin antibodies, as well as blood xylose, iron, and tryglycerides after oral load, were evaluated in 40 celiac children and 43 age-matched patients affected by other gastrointestinal diseases. Each test evaluated individually gave a considerable rate of false-positive and false-negative results. Discriminant coefficients produced for each test were used to compute a score that allowed correct classification of 99% of patients; 2.3% of false-positive and no false-negative results were recorded. This approach improves significantly the overall sensitivity and specificity for celiac disease of these laboratory tests and we propose its use for screening patients to be submitted to jejunal biopsy.

Iron malabsorption in giardiasis.

Among 10 children with giardiasis, eight had iron deficiency; iron deficiency anemia was the main complaint in three. Evaluation of iron absorption by the oral iron load test demonstrated a subnormal response (i.e., increase in serum iron levels of less than 100 micrograms/dl) in all eight patients with iron deficiency. In contrast, in two iron-sufficient patients with giardiasis the response to an oral iron load was normal. Xylose absorption was abnormal in five of the 10 patients. After metronidazole dosing, iron absorption became normal in seven patients but remained abnormal in one patient, who also had IgA deficiency. Xylose absorption became normal in all five patients who underwent a second test, but remained abnormal in the patient with IgA deficiency. Concomitant morphologic-studies of jejunal biopsy material from these patients revealed moderate changes in the intestinal mucosa of two patients. We conclude that malabsorption of iron is a complication of giardiasis.

Sigmoidoscopy, colonoscopy, and radiology in the evaluation of children with rectal bleeding.

The authors evaluated the diagnostic role of sigmoidoscopy, colonoscopy, and double contrast radiology in 103 children with rectal bleeding, with or without other gastrointestinal symptoms. The children's mean age was 44 months, with a range from 1 month to 12 years. In 74.5% of the subjects investigated, visual inspection of the anus and sigmoidoscopy with rectal biopsy disclosed a positive diagnosis. Of the remaining patients, a conclusive diagnosis was reached by either colonoscopy or double contrast radiology in all but six patients. These six, with mild painless hematochezia, remained without a diagnosis. The diagnostic procedure in pediatric patients with rectal bleeding should include an initial visual inspection of the anus, and sigmoidoscopy; air contrast enema and colonoscopy should be performed only in children whose sigmoidoscopy is negative, in diagnostic assessment of inflammatory bowel disease, and in cases of recurrent bleeding after removal of rectal polyps. Colonoscopy is important also in the follow-up examination of children with inflammatory bowel disease and allows the removal of polyps located in the proximal colon.

[Carcinoid cardiopathy]. / La cardiopatia da carcinoide.

A series of 5 males and 2 females aged 23-73 yr with carcinoid cardiopathy is presented, all of them with clinical and instrumental signs of liver metastasis. The main clinical signs were dyspnoea and asthenia rendered ingravescent by effort, and, in the later stage, a frank picture of congestive cardiac decompensation. All subjected presented stethoscopic evidence of tricuspid valvulopathy, combined with pulmonary stenosis in 2 cases. The ECG picture displayed a constant reduction in cardiac potentials, together with right branch bundle block in 3 cases. In cases where an echocardiogram was taken, this confirmed tricuspid involvement. The disease progressed in all cases, and four patients died as a result of terminal liver failure.