The effect of ultrafine WO3 nanoparticles on the organization of thylakoids enriched in photosystem II and energy transfer in photosystem II complexes.

In this work, a new approach to construct self-assembled hybrid systems based on natural PSII-enriched thylakoid membranes (PSII BBY) is demonstrated. Superfine m-WO3 NPs (≈1-2 nm) are introduced into PSII BBY. Transmission electron microscopy (TEM) measurements showed that even the highest concentrations of NPs used did not degrade the PSII BBY membranes. Using atomic force microscopy (AFM), it is shown that the organization of PSII BBY depends strongly on the concentration of NPs applied. This proved that the superfine NPs can easily penetrate the thylakoid membrane and interact with its components. These changes are also related to the modified energy transfer between the external light-harvesting antennas and the PSII reaction center, shown by absorption and fluorescence experiments. The biohybrid system shows stability at pH 6.5, the native operating environment of PSII, so a high rate of O2 evolution is expected. In addition, the light-induced water-splitting process can be further stimulated by the direct interaction of superfine WO3 NPs with the donor and acceptor sides of PSII. The water-splitting activity and stability of this colloidal system are under investigation. RESEARCH HIGHLIGHTS: The phenomenon of the self-organization of a biohybrid system composed of thylakoid membranes enriched in photosystem II and superfine WO3 nanoparticles is studied using AFM and TEM. A strong dependence of the organization of PSII complexes within PSII BBY membranes on the concentration of NPs applied is observed. This observation turns out to be crucial to understand the complexity of the mechanism of the action of WO3 NPs on modifications of energy transfer from external antenna complexes to the PSII reaction center.

Detection of multi-class pesticide residues with surface-enhanced Raman spectroscopy.

The excessive use of pesticides disturbs the natural balance in the environment, creates resistance to pesticides and leads to water and food contamination. Therefore, the implementation of fast, robust and cost effective techniques for the monitoring of pesticides is required. In this work surface-enhanced Raman spectroscopy (SERS) was used for the detection of four common pesticides: atrazine, simazin, irgarol, and diuron. SERS is nowadays considered an effective technique for detection of various analytes in low concentration. Sensitivity of the SERS method depends on the type of substrate that can be either a colloidal solution of metal nanoparticles (NPs) or a metal surface with a suitable nanostructured topology. Here, we have investigated the application of silver nanospheres and silver nanoprisms as SERS substrates in pesticides detection. Colloids with spherical NPs were produced by chemical reduction while Ag nanoprisms were prepared by reducing silver nitrate with borohydride (with citrate as a stabilizing agent) and stirring under a UV lamp for 4 and 10 h. The SERS results have shown that, in the presence of synthesized NPs, it was possible to detect millimolar concentrations of aforementioned pesticides with the exception of diuron.

Proliferative characteristics of Philadelphia-negative myeloproliferative neoplasms - clinical implications.

INTRODUCTION: Philadelphia-negative myeloproliferative neoplasms (Ph- MPN) are characterized by overproduction of one or more blood cell lines. METHODS: We studied the proliferative characteristics of 91 patients with de novo Ph- MPN. Colony-forming cells (CFC) and endogenous colonies (EC), from bone marrow (BM) and/or peripheral blood (PB), were analyzed by colony assay based on methylcellulose. The level of circulating CD34+ cells was determined by flow cytometry. RESULTS: The total number of PB CFC in primary myelofibrosis (PMF) was increased compared to the control sample (P < 0.01) and essential thrombocythemia (ET) (P < 0.05). The highest number of BM and PB EC was observed in polycythemia vera (PV) (P < 0.01). Increased levels of CD34+ cells characterized early-prefibrotic (57%) and advanced-fibrotic PMF (90%) as compared to PV (34%) and ET (32%) (P < 0.01). In the whole Ph- MPN group, the total number of PB CFC (P < 0.01), PB EC (P < 0.05), and CD34+ cells (P < 0.01) correlated with the degree of BM fibrosis. Higher levels of circulating CD34+ cells in PMF correlated with the total number of PB EC (P < 0.05) and degree of BM fibrosis (P < 0.01). CONCLUSIONS: Exploration of the PB proliferative characteristics of Ph- MPN on diagnosis may be helpful in revealing early-prefibrotic PMF. Monitoring the levels of circulating CD34+ cells may provide a sensitive indicator of fibrotic evolution in PV and PMF.

Cytogenetic findings in Serbian patients with Turner's syndrome stigmata.

Cytogenetic findings are reported for 31 female patients with Turner's syndrome. Chromosome studies were made from lymphocyte cultures. Non-mosaicism 45,X was demonstrated in 15 of these patients, whereas only three were apparently mosaic. Eight patients showed non-mosaic and four patients showed mosaic structural aberrations of the X-chromosome. One non-mosaic case displayed a karyotype containing a small marker chromosome. Conventional cytogenetics was supplemented by fluorescence in situ hybridization (FISH) with an X-specific probe to identify the chromosomal origin of the ring and a 1q12-specific DNA probe to identify de novo balanced translocation (1;9) in one patient. To our knowledge, this is the first finding of karyotype 45,X,t(1;9)(cen;cen)/46,X,r(X),t(1;9)(cen;cen) in Turner's syndrome. The same X-specific probe was also used to identify a derivative chromosome in one patient.

Successful treatment of extranodal Hodgkin's lymphoma in a patient with longstanding hairy cell leukemia.

Purine analogs, particularly pentostatin and cladribine, are highly effective in hairy cell leukemia (HCL). Both of these drugs induce responses in approximately 80-95% of patients. However, it is not yet determined if treatment with these drugs can induce second malignancies. Hodgkin's lymphoma is very rare as a second malignancy and there are only 3 reported cases concerning the association of this lymphoma with HCL. We describe a patient with longstanding HCL in complete remission after cladribine, in whom extranodal Hodgkin's lymphoma appeared 8 years after the diagnosis of HCL. Magnetic resonance imaging revealed diffuse intra-osseal neoplastic infiltration of the corpora of the whole spinal column and extra-osseal propagation from the fifth thoracic vertebra into the spinal canal with spinal cord compression. Histological and immunohistochemical analysis of the extradural tumor, which was completely excised, disclosed nodular sclerosis Hodgkin's lymphoma with typical Reed-Sternberg cells that were positive for CD30, CD15, bcl-6, Ki67, p53, EBV LPM-1 and IgG, and negative for CD45, CD20, DBA44, kappa, lambda light chains and IgM. In addition, immunohistochemical analysis of the bone marrow in 1999 showed infiltration with positivity for IgM and negative for kappa light chains and IgG. These findings (expression of different immunoglobulins and light chains on the cells) suggest an independent origin of these 2 B-cell neoplasms. After neurosurgery the patient received 6 courses of the MP-ABVD protocol and achieved a complete remission, which has lasted 16 months thus far.

[Late results of splenectomy in patients with chronic immune thrombocytopenic purpura]. / Udaljeni rezultati splenektomije u bolesnika sa hronicnom imunom trombocitopenijskom purpurom.

Splenectomy is definitive treatment for idiopathic thrombocytopenic purpura (ITP) because it removes both the sites of autoantibody producing cells and also the major site of platelet destruction. The purpose of this study was to evaluate long term results of splenectomised patients with ITP and to determine predictor factors for good response. A 167 patients with chronic ITP (136 females, 31 males), median aged 35 years (17-74) was splenectomised after 2 to 160 months (Median 12) from diagnosis of ITP. Indications for splenectomy were: 6 weeks of steroid therapy with platelet count below 10 x 10(9)/l or 3 months with platelet count under 30 x 10(9)/l, or treatment with prednisone above 30 mg more of 6 months to increase platelet count over 30 x 10(9)/l, or repeated relapses. Postoperative complications developed in 16 patients (9.5%), 3 of them died (1.8%) due to thromboembolism and 17 patients discontinued later controls. During follow up to 172 months (Median 62) 111/147 splenectomised patients were in remission (75.5%), 99 in complete (above 100 x 10(9)/l), 12 in partial (50-100 x 10(9)/l) and 36 patients (24.5%) were relapsed (below 50 x 10(9)/l). Remission was achieved in 79/88 patients (89.8%) with good response to prednisone before splenectomy toward 32/62 patients (51.6%) with poor response to prednisone (p < 0.01). Remission was obtained in 9/11 patients (81.8%) who responded well to intravenous immune globulin (0.4 g/kg x 5 d) and only in 1/8 who did not (p < 0.05). Higher response rate was achieved in patients under 40 years of age (81.6%) than in older ones (63.4%) (p < 0.05). No difference was shown between sex and time intervals (3, 6, 12, 24, 36 or over 36 months) from diagnosis to splenectomy. Splenectomy is an effective treatment of refractory ITP with response rate of 75.5% after median follow up of 62 months. In our patients better results on splenectomy were associated with age less than 40 years, good responses to steroid, and intravenous immune globulin.

[Intraoperative and postoperative complications of splenectomy]. / Intraoperativne i postoperativne komplikacije splenektomije.

Spleen is being surgically removed because of trauma, in diagnostic and-or therapeutical purposes because of the benignant and malignant diseases. The percentage of morbidity during and after splenectomy is relatively low. During surgery might occur bleeding, trauma of the pancreatic tail, stomach, lineal flexure of the colon, left hemidiafragm, left suprarenal gland and upper pole of the left kidney, which must be correspondingly reclaimed during the same intervention. In the early postoperative period, postoperative bleeding, subfrenic abscess, pulmonal atelectasis, bronchopneumonia and left pleural extravasations might occur. Especially is important notification of these events in due time and adequate conservative and surgical treatment. After splenectomy, there is an increase of the number of trombocytes, which might lead to the tromboembolic complications. In the prevention of these complications in the postoperative period prolonged antiagregation therapy is suggested. Postsplenectomy sepsis is very late, general complication of splenectomy, which occurs because of the lower immunity in the child age. To prevent these complications, partial splenectomies, reimplantations of the spleen, prolonged application of the penicillin medicines after splenectomy and antipneumococcal vaccine are performed.

[Successful treatment of primary myelofibrosis with thrombocytosis during pregnancy with alfa-interferon]. / Uspesno lecenje primarne mijelofibroze s trombocitozom u trudnoci primenom alfa-interferona.

Primary myelofibrosis is predominantly a disease of old age, poor prognosis and no curable treatment. Thrombocytosis was observed in only 12% of patients. To our knowledge, there is only one reported case of a young woman with primary myelofibrosis who had a term pregnancy [1]. We report on a 29-year-old woman with thrombocytosis and medical history of two miscarriages in the last 2 years, the iirst at 30 weeks of gestation and the second at 27 weeks. Multiple placental infarctions were observed. She was without symptoms but with moderate splenomegaly 4.5 cm below left costal margin). The platelet count was 651 x 10(9)/L, WBC 7.2 x 10(9)/L with normal differential formula, and haemoglobin level 12 g/dl. Bone marrow biopsy showed fibrotic phase of primary myelofibrosis, with hyperplasia of megacaryocytes, decreased numbers of erythroid and granulocytic cells, and increased amounts of reticulin fibres. Cyctogenetic examination of the bone marrow showed normal female caryotype. Increased numbers of progenitors CFU-Mk, CFU-GM and BFU-E were observed in peripheral blood, and decreased numbers in bone marrow cultures. As the patient wished to become pregnant, the treatment with interferon-a (Roferon A) was started at a dose of 3 MU s.c., three times per week. The platelet count rapidly decreased at a level of 260-370 x 10(9)/L. The pregnancy was diagnosed 5 months later. At the 24 week of pregnancy, platelet count raised to 690 x 10(9)/l and the dose of interferon-a was augmented, 3 MU every day, until delivery. Foetal growth and placental circulation were monitored by serial ultrasonography. At the end of 34 weeks of pregnancy, it was noted that placental flow became insufficient, and after foetal lung maturity was stimulated with dexamethasone, Cesarean section was performed. Male baby was born, weighting 2000 g, with respiratory distress syndrome. This complication was successfully treated, and the child is now one year old, with normal growth and development. The mother is still on therapy with interferon-a, 3 MU, three times a week, and the last blood count was as follows: haemoglobin 10.7 g/dl, WBC 6.1 x 10(9)/L and platelet comt 437 x 10(9)/L. In conclusion, according to the clinical results of interferon-d in thrombocytosis and experimental studies which showed the absence of placental transfer of interferon-d, this therapy could be recommended to women with primary myelofibrosis who wish to have a baby.

[Immunophenotyping of leukemic cells in the diagnosis of hairy cell leukemia]. / Znacaj imunofenotipizacije leukemijskih celija za dijagnostiku leukemije vlasastih (hairy) celija.

INTRODUCTION: It is established that immunophenotyping constitutes a useful method in the diagnosis of hairy cell leukaemia. However, no single marker is specific for hairy cell leukaemia. Two-color-flow cytometry can aid in the diagnosis by showing coexpression of CD11c, HC2 or CD25 with pan B cell markers. Recently, Matutes E. et al. [17] proposed a scoring system of immunophenotypic markers, which could be used to evaluate the diagnosis of hairy cell leukaemia. AIM OF THE STUDY: The aim of the study was to: a) confirm previous observations of immunophenotypic characteristics of hairy cell leukaemia; b) identify antibody combinations of two-color immunofluorescence staining that are most useful in the diagnosis of hairy cell leukaemia; c) examine the value of a scoring system of immunophenotypic markers in the diagnosis of hairy cell leukaemia. METHODS: We analyzed peripheral blood of 46 patients with hairy cell leukaemia using indirect immunofluorescence flow cytometry (EPICS-Coulter) with an extended panel of monoclonal antibodies: CD19, CD2O, CD22, CD24, CD10, HLA-DR, CD11c, CD25, HC2, Slg, kappa and lambda light chains. The diagnosis of hairy cell leukaemia in all patients was made using conventional criteria based on cell morphology, TRAP cytochemistry and bone marrow histology. One fourth of patients were also analyzed using two-color flow cytometry with three antibody combinations as follows: CD19 + CD11c, CD19 + CD25 and CD19 + HC2. RESULTS: Our results showed that hairy cells of our patients had a uniform and unique immunophenotype with expression of the following markers: CD19, CD22, CD2O, CD11c and HLA-DR in 100% of patients, CD24 in 93%, CD25 in 88%, Slg in 82%, HC2 in 67%, CD1O in 50%, kappa light chains in 38% and lambda light chains in 35% of patients (Table 1). The level of detectable circulating hairy cells varied widely, from 2% to 93% of total lymphocytes, and 12 patients (26%) with less than 5% of detectable hairy cells were excluded from analysis. Two-color cytometry showed that antibody combination CD19 + CD11c was coexpressed in 100% of patients, CD19 + CD25 in 78% of patients and CD19 + HC2 in 57% of patients (Table 2). Only patients with 5% or more double colored hairy cells for one antibody combination, were included in the analysis. On the basis of our results of in immunophenotyping of hairy cell leukaemia patients and results of other authors (17), we made our scoring system which considers the reactivity with the following markers: CD19, CD11c, CD25 and HC2. Each marker gives 1 point if positive and 0 point if negative. Score 4 had 83% of patients, score 3 had 14% of patients, score 2 had 3% of patients and no patient had score 1 or 0 (Table 3). CONCLUSION: Our results demonstrated that immunophenotyping with a selective panel of monoclonal antibodies could significantly increase the accuracy in diagnosis of hairy cell leukaemia. Two-color flow cytometry with antibody combination CD19 + CD11c showed coexpression of hairy cells in 100% of our patients. The scoring system for hairy cell leukaemia used in our patients showed that high scores 3 and 4 had 97% of patients.

[Results of treatment in patients with hairy cell leukemia with splenectomy, alpha-interferon and deoxycoformycin]. / Rezultati lecenja bolesnika s leukemijom vlasastih celija primenom splenektomije, alfa-interferona i deoksikoformicina.

INTRODUCTION: The ability of interferon alpha and purine analogues to induce long-lasting remissions in the majority of patients with hairy cell leukaemia has revolutionized the treatment [16, 17, 24, 25], but also provoked dilemma and different opinions about the initial treatment or first line therapy. Splenotomy, the first standard treatment, increases peripheral blood counts but half of the patients require a later systemic therapy because of progressive cytopenia. On the other hand, it has been shown that splenotomy performed after achieving complete remission with interferon alpha or purine analogues contributed to spleen infiltration with hairy cells [37, 38]. PATIENTS AND METHODS: In our study we analysed results of treatment of 44 patients with hairy cell leukaemia who were followed-up for 8 years. Patients were treated with three treatment modalities, mostly successively. Splenotomy++ was performed in 34 patients as a first line therapy (Group I); interferon alpha in 24 patients; in 10 patients as a first line therapy without splenotomy (Group II) and in 14 patients as a second line therapy after splenotomy (Group III). Deoxycoformycin was given to 5 patients as a third line therapy. RESULTS: Our results showed that 20 patients (59 percent) treated only with splenotomy were in haematological remission (Group I). The analysis of prognostic variables demonstrated that at diagnosis patients from Group I, treated only with splenotomy, had at diagnosis higher levels of haemoglobin and thrombocyte counts and a lesser degree of bone marrow (b.m.) infiltration with hairy cells (HCs) than they were found in the other two groups of patients (median Hb = 11.3 g/dL, Plt = 133 x 10(9)/L, HCs in b.m. = 77%). Group II had median Hb = 8.6 g/dl, Plt = 72 x 10(9)/L, HCs in b.m. = 88%, and Group III had median Hb = 9.2 10(9)/L, Plt = 61 x 10(9)/L and HCs in b.m = 90%. Survival curves (Kaplan-Meier method) and Log Rank Statistics showed a significant difference in survival-time (Figure 4) among patients groups (p = 0.0427). Group I had median survival of 270 months; Group II 80 months; and Group III 140 months. Our results demonstrated that different prognostic risk groups existed among hairy cell leukaemia patients and could be identified on the basis of Hb level, Plt count and percent of infiltration of hairy cells into bone marrow. At diagnosis the low risk group of patients (Table 1) had levels of haemoglobin more than 11.3 g/dL; platelet counts more than 72 x 10(9)/L; percent of bone marrow infiltration with hairy cells less than 70%, and expression of lambda type of light chains on leukaemic cells. This group of patients was treated only with splenotomy without systemic therapy. In high risk group of patients the levels of haemoglobin were lower than 8.6 g/dL; platelet counts lower than 60 x 10(9)/L; percent of bone marrow infiltration with hairy cells more than 90% and expression of kappa type of light chains on leukaemic cells. The survival of this group of patients, in spite of initial treatment with systemic therapy with interferon alpha, was shorter. Results of treatment with interferon alpha (Group II and Group III) have demonstrated that the duration of remission in patients who were initially splenectomized and later treated with interferon alpha, was longer (median 31 months) than in patients who were treated with interferon alpha without splenotomy (median duration of remission = 13 months) (Table 3). Results of treatment with deoxycoformycin (Table 4) showed that 3/5 of patients achieved complete remission and 2/5 patients achieved partial remission. Four patients were initially splenectomised and att five patients were treated with interferon alpha before deoxycoformycin. CONCLUSION: On the basis of our results, splenotomy can be recommended as the first-line therapy for low risk patients with hairy cell leukaemia; interferon alpha should be the first-line therapy in high risk patients, and the second-line therapy in l

[Results of treatment of patients with advanced multiple myeloma with the vincristine-adriamycin-dexamethasone protocol]. / Rezultati lecenja bolesnika s napredovalim multipnim mijelomom primenom protokola vinkristin-adriablastin-deksametazon.

UNLABELLED: A very few treatment regimens have shown a benefit in patients with multiple myeloma resistant to conventional melphalan/prednosone therapy or similar combinations. The first "biologically designed" protocol for the treatment of advanced, refractory myeloma was a combination of vincristine, doxorubicin and intermittent high-dose dexamethasone, so called VAD regimen. This report summarizes our experience in VAD regimen in the treatment of advanced, refractory myeloma patients, initially treated with melphalan-based chemotherapy. METHODS: Between July 1989 and July 1995, 27 patients with high-tumour-mass stage (Durie Salmon staging system) of the disease were treated with VAD combination. Clinical characteristics of patients are shown in Table 1. There were 17 pts who never responded (9 pts) of who progressed during induction therapy (8 pts). The second group of 10 pts responded to induction therapy and relapsed. Five pts (four with progressive and one with resistant myeloma) were treated with VAD therapy particularly due to significant extramedullary infiltrates. All pts in this study were initially treated with VMCP induction therapy. Seven of them were additionally treated with ABP combination, and this subgroup of pts was characterized as "resistant to melphalan and doxorubicin". The VAD regimen consisted of four-day continuous infusions of vincristine (0.4 mg per day) and doxorubicin (9 mg/m2 per day) in addition to dexamethasone in a dose of 40 mg for four days, beginning od days 1.9 and 17 of each cycle. The response was defined as a reduction of serum myeloma-protein concentration exceeding 75 per cent, with disappearance of Bence-Jones protein excretion. RESULTS: "Good" response to VAD regimen was achieved in 12 of 27 pts (44 per cent), mainly after three cycles of chemotherapy (Table 2). The tumour reduction occurred rapidly (Table 3), without significant myelosuppression. Response-rate was significantly higher in relapsing myeloma pts than in pts with progressive or resistant disease (chi 2 = 4.2; p < 0.05). Neither previous treatment (Table 2) nor the type or paraprotein concentration, degree of marrow infiltration and cytologic type of plasma cells affected the response to VAD. Among 15 pts with "bad" response to VAD, seven died during first four months of treatment. All pts with significant extramedullary infiltrates failed to respond to VAD (chi 2 = 4.91; p < 0.05). Median survival of all pts was 16 months. In responsive pts remissions were of good quality and survival was significantly longer that that in whom treatment failed (Figure 1, left). In responsive pts the median duration of "plateau-phase" was 11 months (Figure 1, right). In three pts who relapsed after the treatment, reinstitution of VAD regimen restored the "plateau-phase". The most important complications of treatment with VAD combination were infections (8 pts) but, fortunately, serious forms (i.e. pneumonia) were observed only in two pts. DISCUSSION: The antitumour effect of VAD regimen originates from a combined effect of doxorubicin and vincristine continuous infusions and intermittent pulses of high-dose corticosteroids. The rationale for protracted administration of vincristine and doxorubicin was based on long generation time and low growth fraction of plasma cells in most patients, while the use of high-dose dexamethasone was based on well-known dose-depend antimyeloma effect of corticosteroids. Using this chemotherapy schedule, significant prolongation of survival was achieved in our responding patients comparing to patients with VAD-resistant myeloma. The major toxic effect of treatment was infection, which was attributed in part to intensive steroid program. Relapse of the disease could be expected about one year after completion of VAD therapy. Nevertheless, the second "plateau-phase" can be obtained upon reinitiation of VAD (ABSTRACT TRUNCATED).

Megakaryocyte progenitors in paroxysmal nocturnal haemoglobinuria are sensitive to complement.

We have investigated growth in vitro of bone marrow megakaryocytic progenitors (CFU-Mk) in 7 patients with paroxysmal nocturnal haemoglobinuria (PNH) to determine the sensitivity of CFU-Mk to complement. Bone marrow light density mononuclear cells were exposed to fresh or heat-inactivated AB human serum in the presence of medium or isotonic sucrose solution. We found that the proliferative activity of bone marrow CFU-Mk in PNH patients was significantly lower than in controls. In addition, the number of CFU-Mk in PNH bone marrow cells exposed to isotonic sucrose and complement was reduced to 25% of that in PNH cells exposed to isotonic sucrose without complement. In conclusion, our finding showed an increased sensitivity of CFU-Mk in PNH bone marrow cells to complement, supporting the hypothesis that the PNH defect is present at the level of CFU-Mk.

The determination of spontaneous megakaryocyte colony formation is an unequivocal test for discrimination between essential thrombocythaemia and reactive thrombocytosis.

Spontaneous colony formation from bone marrow megakaryocyte progenitors (BMsCFU-Mk) was studied in 24 patients with essential thrombocythaemia (ET), 20 patients with reactive thrombocytosis (RT), 20 patients with polycthaemia rubra vera with thrombocytosis (PRVtr), 16 patients with chronic myeloid leukaemia with thrombocytosis (CMLtr) and 18 normal control subjects (C). The culture medium which was used in the methylcellulose assay in vitro contained 30% of plasma from a single patient with hereditary haemochromatosis. Remarkable BMsCFU-Mk growth was recorded in all patients with ET but in none with RT or in C. BMs-CFU-Mk were present in 11/20 patients with PRVtr and 7/16 patients with CMLtr. Spontaneous bone marrow erythroid progenitors (BMsBFU-E) were also determined in these patients. BMsBFU-E were found in 21/24 patients with ET and none in the patients with RT and C. All patients with PRVtr and one patient with CMLtr showed BMsBFU-E. We conclude that our implementation of the in vitro methylcellulose assay allows the BMsCFU-Mk to be used as an unequivocal test for discrimination between ET and RT which has not been shown in previously published studies. In addition, we present evidence that in 10 patients BMsCFU-Mk and/or BMsBFU-E growth in the test persisted after long-lasting haematological remission.

Three monoclonal IgG components, an IgG4(lambda), an IgG2(kappa) and an IgG1/IgG3 (kappa) Gm(f,b) hybrid, in a single myeloma patient.

An unusual triclonal IgG combination in the serum of a 56-year old male with clinical stage IIIB multiple myeloma is reported. The patient initially had an IgG4(lambda) monoclonal protein in his serum and later developed an IgG2(kappa) and an IgG (kappa) which possessed the characteristics of both IgG1 and IgG3 subclasses with an unusual combination of allotypic markers. Three M-proteins did not share idiotypic determinants. A rare class-switch recombination followed by mutation has been considered as a possible mechanism leading to this combination.

[Chronic lymphocytic leukaemia associated with polycythemia vera]. / Hronicna limfocitna leukemija kod bolesnika s pravom policitemijom.

Simultaneous or sequentional but spontaneous occurzence of polycythaemia vera and chronic lymphocytic leukaemia is very unusual. Moreover, the pathogenesis of these two malignancies has not yet been explained. The authors discribed a 64-year-old man with remarkable mild clinical course of polycytheameia vera associated with chronic lymphocytic leukaemia, lasting more than 5 years. Bone-marrow cell culture revealed spontaneous growth of erythroblast progenitors (BFU-E, CFU-E) and reduced number of haemopoietic progenitorus, mainly due to lymphocyte bone marrow infiltration. The patient plasma selectively inhibited growth of the BFU-E and CFU-E progenitor cells of normal bone-marrow, suggesting that some inhibitor of erythrocytopoiesis influenced supression and/or control of one disease by the other.

Successful treatment of refractory pure red cell aplasia secondary to chronic lymphocytic leukaemia with cyclosporine A: correlation between clinical and in vitro effects.

This report presents the case of a patient with PRCA in CLL where in vitro culture studies correlated well with successful CS-A treatment. Before initiating CS-A therapy, coculture studies showed that T-cells from peripheral blood of the patient suppressed the formation of CFU-E and BFU-E colonies by normal bone marrow cells. Normal erythropoiesis reappeared in the bone marrow of the patient 3 weeks after the start of CS-A therapy. At this time, cocultures demonstrated that peripheral blood T-cells no longer inhibited the growth of normal BFU-E, although there was persistent suppression of CFU-E. Six months later the patient was in stable remission from PRCA on maintenance therapy with CS-A. Moreover, cocultures showed no T-cell inhibition of normal BFU-E or CFU-E colony formation. The strong correlation between in vitro culture studies and a beneficial clinical outcome observed in this case suggests that in vitro cultures could be used to monitor CS-A treatment in patients with PRCA in CLL.

[Leukemic infiltration of gastrointestinal system: unusual complication of hairy cell leukemia]. / Leukemijska infiltracija gastrointestinumskog trakta: neuobicajena pojava u toku leukemije s vlasastim celijama.

Hairy cell leukemia is a rare chronic lymphoproliferative disease, characterized by pancytopenia, splenomegaly and infiltration of the bone marrow and peripheral blood by abnormal lymphoid cells with unique morphology and specific cytochemical staining for the tartarate resistant isoenzyme of acid phosphatase. Leukemic infiltration of the gastrointestinal system is an extremely rare (less than 1%) complication during the course of the hairy cell leukemia. The authors presented a patient in whom the correct diagnosis of this unusual complication was of particular importance for the initiation of alpha interferon therapy. After successful treatment with alpha interferon the patient died, few months later, from unrelated disease-acute anteroseptal myocardial infarction.

Hairy cell leukemia associated with congenital dyserythropoietic anemia type II (HEMPAS).

The case history of a patient with hairy cell leukemia and congenital dyserythropoietic anemia type II (HEMPAS), is described. However, simultaneous occurrence of these two diseases, one of which is hereditary and the other acquired, imposed significant diagnostic problems.

Epidemiology of hepatitis D virus (delta) infection in Yugoslavia.

In 614 HBsAg-positive Yugoslavian patients, radioimmunoassay testing for anti-delta showed the presence of this antibody in serum in 11.2%. Of the patients, 213 belonged to a risk group (i.v. drug users, hemophiliacs, hemodialysed patients and patients with posttransfusion hepatitis); a significant number of these patients (63; 29.6%) were found to have anti-delta. A second group was composed of 401 HBsAg-positive patients from the general population (patients with acute hepatitis B, with fulminant hepatitis B and patients with chronic HBV infection); delta infection was found only in six (1.5%). Immunohistochemical methods failed to demonstrate the delta antigen in the livers of 73 patients with chronic HBV infection. Testing the liver of 36 patients with fulminant hepatitis B for delta antigen demonstrated this reactivity in only one (2.8%) liver sample. Delta antigen was also found in the liver of a female patient who underwent biopsy in 1972. The results of this study suggest the HDV is not endemic in Yugoslavia; however, it is frequently found in patients at risk of blood exposure, primarily i.v. drug users.