[Twenty years of severe aplastic anemia treatment at Department of Hematology, University Department of Medicine, Zagreb University Hospital Center, Zagreb, Croatia]. / Dvadeset godina lijecenja teske aplasticne anemije u Zavodu za hematologiju Klinike za unutrasnje bolesti Klinickog bolnickog centra Zagreb.

Aplastic anemia is a bone marrow disease characterized by marrow aplasia and pancytopenia. Because hematopoietic stem cell transplantation (HSCT) cures severe aplastic anemia (SAA), it is the treatment of choice for younger patients. For many years, antithymocyte globulin (ATG) has been standard immunosuppressive therapy for those aplastic anemia patients that have no HLA matched related donor. ATG significantly improves aplastic anemia outcome, especially when combined with cyclosporine (CSP). The response rate varies from 40% to 70% and long-term survival is comparable with patients receiving marrow transplant. From 1983 until 2006, 46 SAA patients received HLA identical sibling marrow graft. In the same period, 50 patients received standard immunosuppressive therapy combined from horse or rabbit ATG, 6 methyl prednisolone and cyclosporine. Out of 46 transplant patients, 27 received a combination of cyclophosphamide and thoraco-abdominal irradiation. The overall probability of survival for SAA patients that underwent marrow grafting is 51%, and for patients receiving immunosuppressive treatment 20%. We analyzed a cohort of patients receiving treatment after 1990 and found the probability of survival to be 64% for bone marrow transplanted patients and 36% for patients receiving immunosuppression. Infection is the main cause of death in both groups. In conclusion, we documented improving results using ATG in patients with SAA.

Acute promyelocytic leukemia after whole brain irradiation of primary brain lymphoma in an HIV-infected patient.

The occurrence of acute promyelocytic leukemia (APL) in HIV-infected patients has been reported in only five cases. Due to a very small number of reported HIV/APL patients who have been treated with different therapies with the variable outcome, the prognosis of APL in the setting of the HIV-infection is unclear. Here, we report a case of an HIV-patient who developed APL and upon treatment entered a complete remission. A 25-years old male patient was diagnosed with HIV-infection in 1996, but remained untreated. In 2004, the patient was diagnosed with primary central nervous system lymphoma. We treated the patient with antiretroviral therapy and whole-brain irradiation, resulting in complete remission of the lymphoma. In 2006, prompted by a sudden neutropenia, we carried out a set of diagnostic procedures, revealing APL. Induction therapy consisted of standard treatment with all-trans-retinoic-acid (ATRA) and idarubicin. Subsequent cytological and molecular ana?lysis of bone marrow demonstrated complete hematological and molecular remission. Due to the poor general condition, consolidation treatment with ATRA was given in March and April 2007. The last follow-up 14 months later, showed sustained molecular APL remission. In conclusion, we demonstrated that a complete molecular APL remission in an HIV-patient was achieved by using reduced-intensity treatment.

Treatment of relapsed or refractory aggressive non-hodgkin lymphoma with two ifosfamide-based regimens, IMVP and ICE.

We report the outcomes of 45 patients with relapsed or refractory aggressive non-Hodgkin's lymphoma (NHL) treated with a combination of ifosfamide, carboplatinum and etoposide (ICE) and 28 patients treated with a combination of ifosfamide, methotrexate and etoposide (IMVP) during two 5-year periods. The response rate (RR) to ICE was 47%, 2-year overall survival (OS) 31% and 2-year event-free survival (EFS) 22%. These results were similar to those obtained with IMVP (RR 39%, 2-year OS 23%, 2-year EFS 13%; p=0.355 for RR, 0.275 for OS, 0.668 for EFS). Higher IPI scores and refractoriness to treatment were negative prognostic factors, immunophenotype (B vs. T) had no influence on prognosis. Changing from IMVP to ICE does not substantially improve the outcome of patients with relapsed or refractory aggressive NHL. Patients with relapsed/refractory aggressive B-NHL do not have a superior outcome in comparison to those with T-NHL if treated with chemotherapy alone.

Long-term results of conventional-dose salvage chemotherapy in patients with refractory and relapsed Hodgkin's disease (Croatian experience).

BACKGROUND: The aim of this study was to analyze outcome of patients with Hodgkin's disease (HD) in whom first-line chemotherapy with mustine/vincristine/procarbazine/prednisone (MOPP) had failed. PATIENTS AND METHODS: From January 1982 to December 1989 among 210 patients treated with MOPP and radiotherapy to initial bulky sites, 65 patients were primary refractory to or relapsed after initial treatment. RESULTS: Twenty-nine of 65 patients (44%) were primary refractory to initial chemotherapy, 20 relapsed within 12 months after complete remission (CR) and 16 relapsed after CR that lasted more than 12 months. Patients with primary refractory HD and early relapse (<12 months after CR) were treated with doxorubicin/bleomycin/vinblastine/darcarbazine. In patients with late relapse (>12 months after CR) MOPP was repeated. The median follow-up for all patients was 115 months. The overall response rate was 63%. Thirty-three patients (51%) achieved a second CR and eight patients (12%) partial response. Remission rate was greatest in patients with late relapse (CR >12 months) (75 versus 55% for early relapse versus 35% for primary refractory HD) (P <0.01). At 10 years, overall and failure-free survival rates were 21 and 16%, respectively. Patients who were in first remission longer than 12 months had a superior overall survival (37 versus 18% for early relapse) and failure-free survival (24 versus 10% for early relapse). No patient with primary refractory HD was alive beyond 52 months after initial treatment failure (P <0.01). Main prognostic factors were duration of the first remission and tumor bulk at relapse. CONCLUSIONS: Our results confirm previous observations that a significant proportion of patients with HD who experience induction treatment failure cannot be cured with conventional treatment and probably need more aggressive therapy.

Reed-Sternberg cells in the pathogenesis of Hodgkin's disease.

Hodgkin/Reed Stemberg (HRS) cells mediate the classical features of Hodgkin's disease. However, because of their rarity in tumor tissue, little is known about their origin and function. Recent advances in biotechnology, including the single cell manipulation, enabled the insight into the biology of HRS cell. It has been demonstrated that in the great majority of cases they are of germinal center B cell origin, with highly developed interactive network with adjacent cells via expression of cell adhesion molecules, tumor necrosis factor receptor superfamily, and elaboration of different cytokines.

Possibilities and limits of treatment in patients with thrombotic thrombocytopenic purpura.

Results of treatment of 18 patients fulfilling the criteria for TTP are presented. Thrombocytopenia was present in all patients (100%). Sixteen of the 18 patients (88.8%) had mental status changes, and seven of the 18 patients (38.8%) had renal impairment. One patient had a secondary type of TTP, caused by non-Hodgkin's lymphoma of the large intestine (that was diagnosed later) and was excluded from the study. Immunosuppresive therapy with steroids, plasma exchange and replacement of removed volume with fresh frozen plasma in a dosage of 25 ml/kg body weight resulted in a statistically significant increase of platelet count (P = 0.00222), and a significant improvement in consciousness defined by increased GCS after 2 weeks (P = 0.00222). In two patients renal function recovered, and in one of them hemodialysis was no longer needed. This improvement in a small group of patients had no statistical significance. TTP recurred in seven patients. High doses of steroids caused serious side effects in two patients: in one patient, steroid diabetes, and in the other one, intestinal perforation.

Molecular monitoring of minimal residual disease in acute promyelocytic leukemia by the polymerase chain reaction assay for the PML/RAR alpha (retinoic acid receptor-alpha) fusion transcript in patients treated with all-trans retinoic acid followed by chemotherapy.

Five acute promyelocytic leukemia (APL) patients who achieved a complete remission (CR) with all-trans retinoic acid (ATRA) underwent residual disease monitoring through reverse transcription polymerase chain reaction (PCR) for PML/retinoic acid receptor-alpha (PML/RAR alpha) fusion transcript. All received consolidation chemotherapy in CR, one in the form of autologous bone marrow transplantation (ABMT). In four of the patients PCR was positive for the PML/RAR alpha transcript immediately after ATRA treatment and/or after the first consolidation chemotherapy course. In the patient treated with ABMT, positivity was still detected six months after ABMT. One patient given five repeated courses of chemotherapy was PCR negative for PML/RAR alpha after 14 months in CR. Our pilot study confirmed that ATRA is a highly efficient induction therapy for APL in various stages of the disease, but ATRA alone cannot cure the disease. PCR should be considered a fundamental assay for assessing minimal residual disease in CR that will influence further treatment strategies and permit evaluation of treatment results.

Autologous bone marrow transplantation for haematological malignancies--experiences of the centre of Zagreb.

From April 1988 to May 1993, 71 patients (32 acute myelogenous leukaemia [AML], 24 acute lymphoblastic leukaemia [ALL], 7 Hodgkin's disease [HD], 5 non-Hodgkin lymphoma [NHL], 2 neuroblastoma, 1 chronic myelogenous leukaemia [CML]) were treated with myeloablative therapy followed by reinfusion of cryopreserved autologous bone marrow (ABMT). The majority of patients with acute leukaemia were in first complete remission (CR), while 11 AML patients and 9 ALL patients were in advanced stage of the disease (> I CR or relapse). The BM was reinfused without purging. The conditioning regimen for all ALL and proportion of AML patients consisted of cyclophosphamide (CY) 120 mg/kg and fractionated total body irradiation (TB) in a total dose of 12 Gy. 18 AML patients received busulfan 16 mg/kg instead of TBI. Leukaemia-free survival (LFS) for first CR AML patients was 48% at 43 months with the median follow-up of 17 months. Probability of relapse was 44%. LFS for advanced AML was only 9% and the probability of relapse 89%. LFS for first CR ALL patients was 72% at 53 months with the median follow-up of 15 months, while probability of relapse was only 23%. For advanced ALL, LFS was 32% at 33 months and probability of relapse 64%. Probability of toxic death for first CR patients was 11%. We found a predictive value of viability testing and in vitro CFU-GM assay for haematologic recovery after ABMT. We conclude that ABMT with cryopreserved BM is a relatively safe method for consolidation therapy of AL. The results of treatment are encouraging.(ABSTRACT TRUNCATED AT 250 WORDS)

Nucleolar organiser regions and survival in patients with non-Hodgkin's lymphomas classified by the working formulation.

AIMS: To correlate the numbers of silver staining nuclear organiser regions (AgNORs) in non-Hodgkin's lymphoma classified by the working formulation with survival, the first complete remission, and the length of remission. METHODS: Sixty one patients were included in the study. Paraffin wax sections were stained using silver solution to visualise nucleolar organiser regions. The AgNORs were counted in 150 nuclei of each specimen. Data were examined using the Kruskal-Wallis test, multivariate discriminant analysis, and Cox's regression test. Curves were calculated by the method of Kaplan and Meier. RESULTS: Most patients who were alive had low grade lymphoma (p < 0.01). The first complete remission was obtained more frequently in the low and intermediate grade groups (p < 0.05). The longest survival was found in the low grade group (p < 0.001). The mean number of AgNORs differed significantly in all three groups (p < 0.001). This was also true for area of nuclei (p < 0.001) and length of remission (p < 0.05). In a multivariate analysis the numbers of AgNORs were highly predictive for survival, remission, and the length of remission. CONCLUSIONS: The numbers of AgNORs correlated with survival, remission, and the length of remission in patients with non-Hodgkin's lymphoma.

Prostaglandin E2 for prophylaxis of oral mucositis following BMT.

Between October 1988 and December 1990, 60 patients with leukaemia (25 with AML, 19 ALL and 16 CML) undergoing BMT were randomised in a double-blind clinical trial to receive prostaglandin E2 (PGE) (Prostin E2, 0.5 mg per tablet) or placebo for prophylaxis of oral mucositis. Patients had to dissolve tablets in the mouth three times daily starting 7 days before BMT and continuing until 21 days after BMT. The incidence of severe oral mucositis was similar for both groups, 55% in patients receiving PGE and 52% in patients receiving placebo. The duration of severe mucositis did not differ between PGE and placebo groups (chi-square 0.95, p = NS). The incidence of HSV infection was significantly higher in patients receiving PGE. Patients with HSV infection receiving PGE also had a higher incidence of severe oral mucositis. The results presented indicate that PGE is not effective for prophylaxis of oral mucositis in BMT recipients.

Antilymphocyte globulin for treatment of pure red cell aplasia after major ABO incompatible marrow transplant.

A patient developed pure red cell aplasia after ABO incompatible BMT for leukemia. He did not respond to plasma exchange. Antilymphocyte globulin therapy was followed by complete and permanent erythroid recovery with disappearance of recipient-derived isoagglutinins.

AgNORs predictive value of prognosis in non-Hodgkin's lymphoma: comparison with flow cytometric cell cycle analysis.

Paraffin-embedded histopathologic specimens, taken before the commencement of therapy from 14 low-grade and 21 high-grade malignant lymphoma patients, and 9 normal lymph nodes were utilized to analyze six cell DNA-related parameters. The flow cytometry technique was used to determine cell-cycle G0/G1, S and G2/M phases, and silver staining to enumerate nuclear organized regions (AgNORs); nucleus surface area was determined by an image-analyzing system. The six parameters and natural logarithm of cell proportion in the S-phase (LS) were determined according to the histologic tumor type and achievement of the first complete remission (CR). All parameters except cell proportion in G1/M cycle phase differed significantly with respect to histologic cell type, but were not related to the achievement of first CR. Inasmuch as the parameters significantly correlated with each other, multivariate discriminant analysis and proportional hazard regression were applied to estimate their discriminant/predictive values with respect to tumor malignancy. AgNORs proved to be far superior in all three clinical parameters, S-phase was significantly predictive for the achievement of first CR, and LS for tumor histology type. The statistical model applied narrowed down the analysis of seven parameters to two with respect to tumor histology type (AgNORs and LS) and achievement of first CR (AgNORs and S), but only to one for overall patient survival (AgNORs). Only the model for tumor histology type discrimination was statistically significant (R2 = 0.904, p < 0.001). It appears that AgNORs may be of utmost predictive importance for the clinical outcome in NHL.

[The nucleolar organizer region in malignant non-Hodgkin's lymphoma]. / Regija nukleolarnog organizatora u malignih ne-Hodgkinovich limfoma.

Nucleolar organizer region-associated proteins (AgNORs) have been studied in parafin sections of 15 non-Hodgkin's lymphomas by a silver staining technique. On the basis of the Working Formulation of malignant non-Hodgkin's lymphomas all studied cases were divided into three groups. A statistically significant difference was found between number of AgNORs in the nuclei of low-grade lymphomas (mean 2.654 per nucleus), and also between intermediate-grade (mean 1.588 per nucleus) and high-grade malignant lymphomas. Significant difference was also observed between AgNOR area in high-grade and low-grade malignant lymphomas and between high-grade and intermediate-grade malignant lymphomas. However no statistically significant differences between all three studied groups with regard to the size of nucleus were found. It is suggested that this method can be very useful in the diagnosis of malignant non-Hodgkin's lymphomas.

[Sepsis caused by Yersinia enterocolitica in a female patient with liver cirrhosis]. / Sepsa izazvana jersinijom enterokolitikom u bolesnice s cirozom jetre.

As a primarily intestinal pathogen. Yersinia enterocolitica (Y. e.) may cause generalized infection in patients with malignant and other serious diseases or immunodeficient subjects. In certain conditions, elevated serum and tissue iron concentrations represent an additional risk factor for systemic infection with this opportunistic bacterium. In our patient, Y. e. septicemia developed during liver cirrhosis decompensation. Clinical signs of infection were alleviated by appropriate antibiotic therapy (gentamycin, cefuroxime), but as septicemia had been present for several days prior to therapy, it aggravated the patient's general condition, which entailed the development of hepatorenal syndrome and eventually lethal outcome.