[Morphological studies of erythrocytes in congenital dyserythropoietic anemia type II]. / Badania morfologiczne erytrocytów we wrodzonej anemii dyserytropoetycznej typ II.

The etiology of congenital dyserythropoietic anemia (CDA) type II is unknown. The diagnosis is based on morphologic and immunologic criteria. We present three girls with well documented CDA II who were followed for 5-8 years. The anemia was mild, progressive body iron overload was found. In none of the girls splenectomy was indicated. Morphologic features of ++erythrocytes and bone marrow erythroid cells were studied by means of light and electron microscopy. Up to 45% of erythrocytes showed invaginations with endocytic cisterns and shape abnormalities (echinocytes, anisocytosis, microcytosis). Typical abnormalities of the external surface of RBC membrane: invaginations, depressions, pits and plaques were shown in the scanning electron microscopic studies. Our studies indicate that the morphological features of erythrocyte in our patients may be consequence of the biochemical changes in the membranes and may contribute to the shortened life span of erythrocytes in patients with CDA II.

Lack of interleukin-2 (IL-2) dependent growth of TAC positive T-ALL/NHL cells is due to the expression of only low affinity receptors for IL-2.

Binding of interleukin-2 (IL-2) to high affinity receptors on activated normal T cells was shown to be the essential step in induction of proliferation of such cells. The finding of abundant IL-2 receptors on malignant T cells in adult T cell leukemia suggested a deregulation of the IL-2/IL-2 receptor system and was assumed to account for aberrant growth in malignant disorders of T cells. In this study we use malignant T cells from nine patients with the clinical diagnosis of T-ALL or T-NHL and did not detect IL-2 dependent growth under conditions in which normal T cells responded to IL-2. IL-2 receptors comparable in numbers to activated T cells were found on T-ALL/T-NHL cells stimulated with PHA and PMA. However, binding studies using radiolabeled IL-2 indicated that the receptors present on malignant T cells were not able to bind to IL-2 with high affinity. Therefore, if IL-2 is involved in the proliferation of malignant T cells, its mechanism of growth regulation may be different from the one for normal T cells. Alternatively, IL-2 may not play a role in the regulation of growth of malignant T cells in vitro.

New aspects in the treatment of childhood rhabdomyosarcoma: results of the German Cooperative Soft-Tissue Sarcoma Study (CWS-81).

The results of the German Co-operative Soft-Tissue Sarcoma Study (CWS-81) of the treatment of rhabdomyosarcoma are presented. Prior to the introduction of chemotherapy only 10%-20% of the children were successfully treated. Combined multi-agent cytostatic treatment improved the results dramatically. In patients with primary stage III rhabdomyosarcoma, local tumour control by surgery or radiotherapy should be undertaken earlier than week 16, if complete remission has not been achieved by 7-9 weeks. Patients with complete remission or partial tumour regression should be treated with the same combination of chemotherapy, while only partial responders need radiotherapy in addition. Patients with tumours which are primarily resectable without mutilation have a 90% chance of cure; this also applies to patients with primarily unresectable tumours who achieve complete remission after 7-9 weeks of chemotherapy. Total disease-free survival rate for stage III rhabdomyosarcoma patients was 53%. The role of surgery includes primary removal of the tumour or assessment of remission by means of histological spot checks.

[Results of treatment of primary metastatic Ewing sarcoma. A retrospective analysis of 48 patients]. / Behandlungsergebnisse beim primär metastasierten Ewing-Sarkom. Eine retrospektive Analyse von 48 Patienten.

The clinical presentation of the disease and the results of treatment in 48 patients with metastases at diagnosis of Ewing's sarcoma, entered into the Cooperative Ewing's Sarcoma Studies (CESS) 1981 and 1986 of the German Society of Pediatric Oncology (GPO), were analysed. The period of observation ranged from 1 to 82 months, the median relapse-free time was 26 months. There was a male predominance of 35 to 13, which was even more pronounced in patients older than 15 years. The predominant localization of the primary tumor was the pelvic region, followed by the extremities, the chest wall, and the spine. The most common site of primary metastases were the lungs, followed by bone and bone marrow. Nine patients presented with combined metastases. The disease-free survival according to Kaplan-Meier life-table analysis was 18% after 7 years. Best results were obtained in patients with pulmonary metastases only, who underwent surgical resection of the primary tumor and received radiation to the lungs. Without bone marrow transplantation the prognosis of patients with bone metastases was extremely poor.

Malignant peripheral neuroectodermal tumors. A retrospective analysis of 42 patients.

The clinical presentation of the disease and the results of treatment in 42 patients with malignant peripheral neuroectodermal tumors (MPNT) entered into the Cooperative Ewing's, soft tissue, and neuroblastoma trials of the German Society of Pediatric Oncology were retrospectively analyzed. Within the Ewing's sarcoma trial, patients with chest wall lesions were particularly analysed for MPNT features. The period of observation ranged from 15 to 86 months; the median relapse-free time was 24 months. There were 28 male and 14 female patients, the median age of patients was 15 years (range, 9 months-23 years). Thirty-two patients had localized disease (M0), and ten patients presented with primary metastases (M1). The predominant location of the tumors was the thoracopulmonary region, followed by the extremities, the abdominal/pelvic, and head and neck region. Thirty-one of 42 tumors involved the adjacent bone. The disease-free survival according to Kaplan-Meier life-table analysis was 56% +/- 11% for Stage M0 patients at 3 years. Nine of ten patients with M1 disease showed progression of their disease. Most patients had combined modality treatment with surgery, chemotherapy and radiation therapy. Best results were obtained with extensive surgery. Radiation doses ranged from 20 to 60 Gy and could not be correlated with the outcome of the disease. Most recurrences occurred at the site of the primary tumor. In patients with primary chemotherapy after biopsy-proven diagnosis, the responsiveness of this disease to chemotherapy could be demonstrated. Combination chemotherapy containing anthracyclines and high doses of alkylating agents appeared superior.

Dihydrofolate reductase-activity in brain tissue. Effect of X-irradiation.

The mechanism responsible for the toxic late effects of cranial irradiation, followed by the administration of systemic methotrexate (MTX) on brain tissue, is still under discussion. We studied the influence of X-irradiation on dihydrofolate reductase (E.C. 1.5.1.3) activity (DHFR), the enzyme inhibited by MTX. New Zealand white rabbits, 6-9 weeks old, underwent 24 Gy fractionated or 20 Gy single-dose brain irradiation using a 60Co source. Before, immediately following, and 1, 2, 4, 12 weeks after irradiation, DHFR was measured in brain and liver tissue by a photometric assay. DHFR in brain tissue was 11.9 +/- 2.9 mU/g wet weight (ww) X h and in liver tissue 121.8 +/- 24.2 mU/g ww X h. Fractionated brain irradiation with 2 Gy per day produced no significant changes in brain DHFR. Single-dose cranial irradiation significantly decreased brain DFHR (7.3 +/- 0.6 mU/g ww X h). Suppression of the developmental increase of DHFR by X-irradiation in young rabbits could be excluded by determining the unchanged brain-to-liver ratios of DHFR in the animals with fractionated brain irradiation.

Synaptophysin identified in metastases of neuroendocrine tumors by immunocytochemistry and immunoblotting.

Synaptophysin, an Mr 38,000 integral membrane glycoprotein of neurotransmitter vesicles, has been identified in diverse primary neuroendocrine (NE) tumors of both neural and epithelial origin (Wiedenmann and co-workers, Proc Natl Acad Sci USA 1986; 83: 3500-3504). In the present study, metastases of several types of NE tumors, including medullary thyroid carcinoma, gastrinoma, insulinoma, small (oat) cell carcinoma of the lung, gastrointestinal carcinoid, and neuroblastoma, were examined for the presence of synaptophysin by immunocytochemistry, with the use of tissue sections as well as centrifuged cell suspensions and by immunoblotting of tumor proteins. The results show that expression of synaptophysin can be maintained during formation of metastases. Therefore, the authors propose that synaptophysin antibodies be used for the positive identification of metastatic NE tumors, notably in differential diagnosis. The possible implications of these findings for tumor diagnosis are discussed.

[Treatment of acute lymphoblastic leukemia in childhood and adolescence: results of the multicenter therapy study ALL-BFM 81]. / Die Behandlung der akuten lymphoblastischen Leukämie im Kindes- und Jugendalter: Ergebnisse der multizentrischen Therapiestudie ALL-BFM 81.

In therapy study ALL-BFM 81 633 previously untreated patients with acute lymphoblastic leukemia (ALL) less than 18 years of age have been recruited from April 1, 1981 to September 30, 1983 and treated in 37 institutions throughout West-Germany and Austria. Here only therapy results of 611 patients with non-B-ALL are presented. Patients with ALL of B-type are described elsewhere. In this fourth consecutive trial of the BFM study group three major questions have been asked: 1. Is it possible to assess the individual risk for relapse more accurately by the use of a risk factor rather than by the risk score which was the discriminator in studies ALL-BFM 76 and ALL-BFM 79? Does this risk factor discriminate more precisely patients at the highest risk for relapse? Offers more intensive risk-adapted therapy to this patient group a better chance for disease-free survival? 2. In patients at a standard risk for relapse with a risk factor below 1.2--approximately 60% of patients with non-B-ALL--can radiotherapy for prevention of CNS disease effectively be replaced by chemotherapy (intermediate dose Methotrexate)? 3. It is possible to reduce duration of maintenance therapy by 6 months to a total duration of 18 months with no unfavorable effect? To assess the radiation problem in standard risk patients and to evaluate the importance of duration of maintenance therapy two randomisations have been utilized. After a median duration of study ALL-BFM 81 of 4 1/2 years and 3 1/4 years after the study had been closed (date of evaluation January 1, 87) the answers are as follows: 1. For the majority of patients risk-adapted therapy had a curative effect. The probability for event-free survival (EFS) in standard risk patients in slightly above 70%, in medium risk patients 67%. In high-risk patients risk-adapted therapy did not improve prognosis, the EFS being still in the order of 50%. A good assessment of the individual risk for relapse is possible by the newly introduced risk factor. This principle is superior to the risk score used in former studies ALL-BFM 76 and ALL-BFM 79 because a low risk group (risk factor below 0.8) could be identified including approximately 25% of all patients with non-B ALL. Selection, quality, and timing of therapy elements remain the decisive prognostic factors, however. 2. Standard risk patients with a risk factor below 0.8 can effectively be protected for CNS relapse by treatment with intermediate dose Methotrexate.(ABSTRACT TRUNCATED AT 400 WORDS)

[Malignant peripheral neuroepithelial tumors in childhood]. / Maligne periphere neuroepitheliale Tumoren im Kindesalter.

The different therapy modalities and course of disease of 42 patients with a malignant peripheral neuroepithelial tumor are retrospectively analyzed. Therapy was completed in 31 children, 25 of whom had a primary localized tumor and 6 a disseminated neuroepithelioma. 17 of the children with a localized illness survive disease free in contrast to no survivor in the group with a disseminated tumor. The effective chemotherapy combining vincristine, adriamycin, ifosfamide and actinomycin D must be complemented by an efficacious local control because malignant peripheral neuroepithelioma tend to recur locally. The prospective analysis of newly diagnosed patients and a standardized therapy regimen will show if malignant peripheral neuroepithelioma represents a distinct tumor entity, different from a Ewings-Sarcoma.

Clitoromegaly and abdominal tumour as leading signs of a mediastinal non-Hodgkin lymphoma.

A 2-year-old girl was admitted with clitoromegaly, mediastinal tumour and enlarged kidneys. A seventh cranial nerve palsy was associated with an increased cell count in cerebrospinal fluid. The diagnosis of a lymphoblastic lymphoma was confirmed by a cervical lymph-node biopsy. Six years after diagnosis and treatment according to the BFM protocol the girl remains in first complete remission.

[Local recurrence of mediastinal non-Hodgkin lymphomas in childhood]. / Lokale Rezidive bei mediastinalen Non-Hodgkin-Lymphomen im Kindesalter.

Local relapses in mediastinal non-Hodgkin's lymphomas in childhood are rare. Of the 15 children treated in our hospital during the last 11 years, 2 had early local or pleural relapses. At presentation both had extensive pleural effusions, and after completion of induction therapy still some fluid in the pleural space was present. A local irradiation was not performed initially. Be review of the literature for risks and benefit of local irradiation in mediastinal non-Hodgkin's lymphoma, local irradiation in children receiving aggressive chemotherapy should only be considered in few patients; preferentially in those, with incomplete disappearance of the mediastinal tumor or, as we may conclude from our patients, incomplete disappearance of pleural effusion.

[Changes in the blood-cerebrospinal fluid barrier for serum proteins in children with acute lymphatic leukemia]. / Veränderungen der Blut-Liquorschranke für Serumproteine bei Kindern mit akuter lymphatischer Leukämie.

The blood-CSF barrier inhibits permeation of most chemotherapeutic agents into the central nervous system (CNS). The influence of systemic chemotherapy and prohylactic CNS irradiation on the permeability of the blood-CSF barrier was studied in 49 children treated for acute lymphoblastic leukemia (ALL) or non-Hodgkin's lymphoma. To study the permeability of the blood-CSF barrier under treatment according to BFM-ALL protocols, nephelometric determinations of albumin, immunoglobulin G (IgG), and alpha-2-macroglobulin in serum and CSF and total protein in CSF were performed at several time intervals during chemotherapy and prophylactic cranial irradiation. During systemic induction chemotherapy, no significant changes of blood-CSF barrier could be observed. In contrast, in the course of prophylactic CNS irradiation and intrathecal methotrexate application, a significant elevation of albumin, alpha-2-macroglobulin and total protein in CSF, and a significant decrease of blood:CSF ratios for albumin and alpha-2-macroglobulin were observed. IgG did not change significantly. After prophylactic CNS treatment and during maintenance chemotherapy protein concentrations and blood:CSF ratios gradually returned to normal range. This normalization was accelerated by cortisone treatment during the reinduction period.

Changes in the EEG background activity of children with acute lymphoblastic leukemia during cytotoxic therapy.

Thirteen children with acute lymphoblastic leukemia (ALL) were investigated before and during cytotoxic therapy. EEG findings were correlated with the clinical course and the therapy protocol and compared with normal data obtained from 295 healthy children. Frequency analysis of the background activity of the EEG revealed an initial slowing of the background activity prior to therapy and further slowing each time a combination of vincristine (VCR), daunorubicin (DAU) or adriblastine (ADR), prednisone (PRED), and L-asparaginase (L-ASP) was administered. The slowing of the background activity correlated only with the administration of these drugs. DAU, ADR, and PRED are not known to influence the EEG; therefore, VCR and L-ASP remain the primary candidates responsible for the central nervous system alteration.

Effects of local irradiation and i.v. methotrexate on brain morphology in rabbits: early changes.

Brain damage following cranial radiation therapy can be crippling or even life-threatening and has been studied in both patients and animals. An additional toxic effect of chemotherapy has been found in children, who died following brain irradiation and systemic chemotherapy for the treatment of acute lymphoblastic leukemia. To study the interaction of radiation and drugs on brain tissue, we treated rabbits with brain irradiation and/or i.v. methotrexate. For a period of up to 3 months following radiation therapy, brain morphology was compared in seven treatment groups. Weekly doses of methotrexate administered i.v. produced no brain damage. Histological examination showed myelin swelling and beading 14 weeks after fractionated brain irradiation with 24 Gy. Combination of brain irradiation and methotrexate produced additional hypertrophy of microglia and pyknosis of adventitial cells. In none of these groups, even after doses of 48 Gy brain irradiation, was calcification or brain necrosis observed during the first 14 weeks following irradiation.

[Scintigraphic diagnosis of neuroblastoma using meta-iodobenzylguanidine]. / Szintigraphische Diagnostik des Neuroblastoms mit meta-Jod-Benzylguanidin.

In 1979/80 meta-iodobenzylguanidine (MIBG) was introduced as a radiopharmaceutical agent with high affinity to the adrenal medulla. It could be shown, that scintigraphic imaging with 131J labeled MIBG is a sensitive and highly specific method for localization of pheochromocytoma. In this connection we could demonstrate in 1983 that neuroblastoma can be visualized scintigraphically with MIBG as well. Until now 13 patients were examined with 131J-MIBG by our group: In 10 children with histologically proven neuroblastoma we found a specific enrichment in the tumor area. Besides the primary tumor local recurrence and metastases to bones, bone marrow and brain were detected. There was no neuroblastoma manifestation without MIBG uptake. In 3 patients with tumors other than neuroblastoma no uptake of MIBG was noticed. The results agree with those of other studies: If the physiological pattern of distribution is taken into consideration and if the proper imaging technique is adapted, 131J-MIBG scintigraphy is a highly sensitive and specific method for staging, monitoring of disease and follow-up of neuroblastoma.

Determinants of plasma arginine-vasopressin in children.

Plasma concentration of arginine-vasopressin (AVP) was measured in 145 healthy subjects aged one day to 18 years of age. AVP decreased with age immediately after birth. Above one year of age values of children did not significantly differ from those in adults. AVP significantly correlated with plasma and urine osmolality after water deprivation during 16 h. Plasma AVP rose during exercise and fell after volume expansion. Nausea and vomiting are potent nonosmotic determinants of AVP release in children. Nonosmotic factors of AVP release should be controlled when sampling blood for measuring AVP in children for diagnostic and investigational purposes.

Effects of methotrexate on rabbit testes. Part 1: Morphological changes.

Methotrexate (MTX) has the potential of eradicating small infiltrations of leukemic cells in the gonads. We examined the morphological changes in rabbit testes after a single dose (57.5 mg/kgBW) or repeated low doses (6 mg/kgBW once a week for 14 weeks) of MTX IV. Fertility rate and spermatogenic activity were evaluated using the tubular fertility index (TFI). Testicular MTX concentrations (measured by RIA) were in the same range in both groups. Only after repeated MTX application were reduction of TFI and signs of germinal cell line degeneration found. When given repetitively, MTX therapy may modify the fertility and tubular morphology. Long-term follow-up will show how these changes affect future fertility.

Effects of methotrexate on rabbit testes. Part 2: Hormonal changes.

Thirty mature and peripubertal male rabbits were examined for endocrine function and tissue methotrexate (MTX) concentration after single (57.5 mg/kgBW, group 1) and repeated (6 mg/kgBW, once a week for 14 weeks, group 2) MTX doses. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and androstenedione plasma levels were measured by radioimmunoassay (RIA). We found elevated plasma FSH levels in both groups. Elevated plasma androstenedione level and reduced plasma testosterone level in group 2 suggest an enzymatic defect in the gonadal steroid synthesis. Unchanged LH plasma level, when compared to controls, is thought to be the result of a combined effect of MTX on the gonadal steroids and gonadotropin synthesis.

[Effect of a local surgical procedure on the incidence of metastases following neoadjuvant chemotherapy of osteosarcoma]. / Einfluss des lokalchirurgischen Vorgehens auf die Inzidenz von Metastasen nach neoadjuvanter Chemotherapie des Osteosarkoms.

Following preoperative chemotherapy of 9-18 weeks duration limb salvage procedures were performed instead of ablative surgery in about 1/2 of the patients (pts). Overall continuous disease-free survival rate is 69% (80/115) at 37 (21-51) months. 5 pts died from therapy related complications, 4 developed a local failure (2 following amputation and 2 following limb salvage each) and 26 pts developed pulmonary metastases. The incidence of pulmonary metastases after en bloc resection, but not after shank rotation plasty, was found to be significantly increased over that after ablative surgery (83% vs 60% metastases free survival (MFS) at 40 months, p less than 0.05). The outcome was most unfavourable following en bloc resection of large tumors (36% MFS) and of tumors poorly responding to preoperative chemotherapy. Delaying surgery for preoperative chemotherapy in itself did not influence MFS-rate but it enabled a thorough planing and preparation of surgical procedures. Chemotherapy has very much improved the prognosis of osteosarcoma, trials on limb salvage surgery are indicated therefore. However, these procedures appear to be hazardous by increasing the rate of pulmonary metastases. Until the underlying mechanisms are not uncovered and preventive strategies worked out, limb salvage surgery in osteosarcoma has to be regarded and handled as an experimental procedure.

[Bone tumors and tumor-like changes in the neonatal period and in infancy]. / Knochentumoren und tumorähnliche Veränderungen im Neugeborenen- und Säuglingsalter.

Bone tumours and tumour-like lesions occurring in newborns and infants are described presenting 5 own cases and a literature review. Age specific features in diagnosis, therapy and prognosis are discussed. Due to the rarity of these diseases a centralized registry at a national level is recommended.