[Assessment of Heavy/ Light Chain Pairs of Immunoglobulin (Hevylite assay) -  Benefit for Stratification of Multiple Myeloma?]. / Vysetrení páru tezkých/ lehkých retezcu imunoglobulinu (Hevylite assay) -  prínos pro stratifikaci mnohocetného myelomu?

BACKGROUND: The aim of the study was the comparison of two novel stratification models in multiple myeloma (MM), ie. according to Avet- Loiseau (A L) and according to Ludwig (L), based on the HLC r index (ratio of serum levels of involved- HLC/ uninvolved HLC, ie. HLC κ/ HLC  λ assessed using ie. nephelometric/turbidimetric technique using specific polyclonal antibodies on a Binding Site SPA(PLUS)) technique) and ß(2) microglobulin (ß(2) M) with selected prognostic factors (PF) of MM and staging systems according to Durie- Salmon (D S) and International Staging System (ISS). PATIENTS AND METHODS: In a cohort of 132 patients (94 with IgG and 38 with IgA type of MM) at the time of dia-gnosis, we assessed HLC r, select-ed PF and D S, ISS, A L and L stratification systems. RESULTS: Unlike in IgA isotype, in IgG isotype we found a significant relationship of HLC r to stratification according to D S and ISS with the difference between A and B substages according to D S (p = 0.049) and between ISS stages 1 vs. 3 (p = 0.001). In the IgG group, there was highly significant relationship of the depth of Hb and albumin decrease and ß(2) M increase to the results of stratification according to ISS, A L and L model (p < 0.0001), increase of LDH in the ISS system and A L, and creatinine according to ISS and L but not the relationship of the stages according to any of the stratification systems to the values of FLC r (ratio of serum free light chains κ/ λ of immunoglobulin), thrombocytes and Ca. In the IgA type, there was a significant relationship of the depth of the decrease of Hb, thrombocytes, albumin and increase of ß(2) M to the results of stratification according to ISS, A L and L and increase of creatinine in the case of ISS, but not of the values of FLC r, Ca and LDH in the case of any of the stratification systems. The degree of correlation of selected PF, especially of Hb, albumin and ß(2) M, event. of thrombocytes, LDH and creatinine to the stages according to ISS and to stage 1-3 according to A L and L model was in IgG vs IgA isotype significantly different (p < 0.0001- 0.030). Staging system according to ISS had proportional distribution of stages 1- 3, whereas in the A L model prevailed in IgA and IgG isotype risk category 2, ie. intermediate-risk (47.3 and 44.7%) and in the L model prevailed risk category 3, ie. high-risk (41.5 and 52.6%) with low count of category 1, ie. low- risk category (23.4 and 10.5%). McNemar- Bowker test of symmetry showed in both types of MM the highest concordance between the stratification according to D S and L in category 3, ie. high-risk (31.9 vs. 28.9%) with overall accord only in 53.2 and 42.1% and with significant shift in the case of IgG isotype only (p = 0.036). In IgG and IgA isotype there was an overall concordance in the distribution of categories 1- 3 according to ISS vs. A L (62.4 and 63.2%) but with significant shift of the stratification (p = 0.002 and 0.028). In the case of IgG and IgA isotype there was a close relationship between the models A L and L (64.5 and 81.6%) with significant stratification shift (p < 0.0001 and 0.030). CONCLUSION: The new stratification models for MM according to A L and L are easily practically applicable, with close relationship to principal PF but they need separate assessment of IgG and IgA isotypes of MM. The choice of optimal model for routine practice needs a validation study aimed at progression free survival and overall survival.

[Attainment of complete hematological remission is crucial for extended survival of AL amyloidosis patients with cardiac involvement]. / Dosazení kompletní hematologické remise je zásadní pro prodlouzení prezití nemocných s AL amyloidózou a srdecním postizením.

BACKGROUND: Symptomatic cardiac involvement is the most important prognostic factor in AL amyloidosis patients. Longterm survival is limited not only by cardiac involvement condition, but also by limited choice of treatment with unsatisfactory results. The aim of the present report is to assess the effect of achieved treatment response on survival of AL amyloidosis patients with symptomatic cardiac involvement under conventional treatment. MATERIAL AND METHODS: The monitored patient set consisted of 19 patients with systemic AL amyloidosis and symptomatic cardiac involvement, treated and monitored at the III. Clinic of Internal Medicine between 2004 and 2012. The male : female ratio was 17 : 2, and the age median was 64 (range 48 to 78 years). Thirteen patients died within the monitored period. Functional status was defined according to the NYHA classification, where five patients had class II involvement, 10 patients had class III involvement, and four patients had class IV involvement. Treatment response was assessed by the application of modified IMWG and ISA criteria; all patients were undergoing conventional treatment. Nine patients were treated by a combination of alkylating agents (alkeran, cyclophosphamide), six were treated by a combination treatment with thalidomide, and four were treated by a combination of bortezomib and dexamethasone. Data were analyzed with software SPSS v. 15 (SPSS, Inc., Chicago, USA). Log Rank Test was applied to survival evaluation. RESULTS: The statistical analysis included only 13 patients who underwent at least three months of treatment, where six patients attained complete remission (CR), four patients attained partial remission (PR), and three patients attained only stabilization of disease (SD). Significant difference in patient survival was found to be correlated with attained hematological response, where the patients who attained CR had median survival of 39 months vs 10 months in patients who attained PR or SD (p = 0.005). CONCLUSION: The results indicate that attainment of complete hematological remission is associated with significantly longer survival of AL amyloidosis patients with symptomatic cardiac involvement.

[The importance of cardiac bio-marker assay for the stratification and monitoring of AL amyloidosis patients -  single center experience]. / Význam stanovení kardiálních bio-markeru ve stratifikaci a sledování nemocných s AL amyloidózou -  zkusenosti jednoho centra.

INTRODUCTION: Cardiac involvement is a dominant prognostic factor in AL amyloidosis patients. A detailed assessment of the presence and degree of cardiac involvement utilizes an array of noninvasive investigation methods, particularly echocardiography and MRI; laboratory parameters include troponins and natriuretic peptides. Cardiac involvement detection aside, cardiac bio-markers are used as a relatively strong stratification and prognostic factor. OBJECTIVE: The presentation of cardiac bio-markers assay applications in AL amyloidosis patients at an individual treatment center. PATIENTS AND METHODS: The monitored patient set consisted of 22 patients with histologically confirmed AL amyloidosis, of whom 18 met the criteria for cardiac involvement. Levels of cardiac bio-markers troponin T (TnT) and Nterminal probrain natriuretic peptide (NT ProBNP) were determined in all patients. Risk stratification of the patients utilized the Mayo staging system which is based on both bio-markers assays; Log Rank Test was applied to survival evaluation. RESULTS: Median survival of patients with cardiac involvement stigmata was 10 months vs 60 months survival of patients without signs of cardiac involvement (p = 0.133). Of the 4 patients without cardiac involvement, 1 has shown positive levels of TnT and 2 positive levels of NT ProBNP. All cardiac involvement patients exhibited abnormal levels of NT ProBNP (median 4,752 ng/ l; 415.7- 35,000) as well as positive levels of TnT (median 0.0815 µg/ l; 0.02- 0.986). The application of the Mayo stratification system to the set had determined 2 patients at stage I, 5 patients at stage II and 15 patients at stage III. The median survival of the Mayo I + II group vs the Mayo III group was 60 vs 6 months (p = 0.015), revealing extremely limited survival of stage III patients. Assessment of TnT and NT ProBNP levels relative to treatment response shows that the degree of decrease in both markers depends on maximum treatment response -  respectively the attainment of a complete hematological remission. CONCLUSION: The results, although obtained from a limited set of patients, confirm a definitive benefit of the application of cardiac bio-markers assay in the diagnostic and therapeutic algorithm of AL amyloidosis patients. The Mayo stratification system utilizing the cardiac indicator values represents a robust tool for risk stratification of AL amyloidosis patients.

Low molecular weight heparins for thromboprophylaxis during induction chemotherapy in patients with multiple myeloma.

BACKGROUNDS: Patients with multiple myeloma have a high risk of venous thromboembolism (VTE), especially during the induction chemotherapy. The aim of our observational study was to determine the impact of prophylaxis with low molecular weight heparin (LMWH) on the incidence of thromboembolic complications. PATIENTS AND METHODS: We analyzed the incidence of thromboembolic events in 258 patients treated with induction chemotherapy containing vincristin, doxorubicin or idarubicin, and dexamethasone, followed by stimulation chemotherapy with cyclophosphamide and G-CSF, and high-dose chemotherapy with melphalan. Two groups of these patients were compared based on the practice of thromboprophylaxis. Patients in the first group (Control, n = 140) were either not treated or treated with a short duration of anticoagulation therapy while the patients in the second group (Prophylactic, n = 118) underwent standard prophylaxis with LMWH throughout the entire period of induction chemotherapy. A total of 102 patients were selected for a close monitoring of the prophylactic effect of different LMWH doses and to be compared to patients without treatment. RESULTS: Standard prophylaxis with LMWH significantly (p < 0.007) lowered a risk of VTE when compared to patients without such prophylaxis (3.4% versus 12.9%, respectively). Furthermore, analysis of the subgroup of 102 patients revealed that higher LMWH doses (> 70 IU/kg per day) achieved full prophylaxis in 28 patients while lower doses were less effective leading to DVT in 3 (7.7%) out of 39 patients. In contrast, VTE was diagnosed in 5 (14.3%) out of 35 patients without any LMWH prophylaxis. CONCLUSION: Prophylaxis with LMWH leads to a significant reduction of the risk of thromboembolic complications during the induction chemotherapy in patients suffering from MM. The prophylactic effect of LMWH is dose-dependent.

Analysis of the serum levels of selected biological parameters in monoclonal gammopathy of undetermined significance and different stages of multiple myeloma.

The aim of the study was to analyze differences in the serum levels of 8 selected biological parameters between monoclonal gammopathy of undetermined significance (MGUS) and different stages of multiple myeloma (MM), potentially beneficial for distinguishing between the two conditions. The analyzed group of 131 subjects comprised 62 individuals with MGUS and 69 MM patients examined at the time of diagnosis. The serum levels were determined by a quantitative immunoradiometric assay (insulin-like growth factor 1, IGF-1) and quantitative sandwich enzyme immunoassay (osteopontin, OPN; endostatin, ES; macrophage inflammatory proteins 1α/ß, MIP-1α/ß; angiogenin, ANG; and interleukin 17, IL-17). The analysis showed a statistically significant difference in serum concentrations between MGUS and the symptomatic form of MM using the Durie-Salmon (D-S) staging system only in the cases of OPN and stages II and III (0.001 and MM. More benefit may be expected from analyses using multiparametric immunophenotyping of plasma cells and molecular biology methods including gene expression analysis and proteomics.

[Multiple spinal myeloma and its surgical management]. / Mnohocetný myelom pátere a jeho chirurgická lécba.

INTRODUCTION AND AIM: The management of spinal multiple myeloma (MM) is a complex process, including causal treatment (i.e. efforts to suppress the tumor clone), as well as supportive therapy, including surgery. The aim of this article is to present retrospective evaluation of surgical indications in patients with MM or solitary spinal plasmocytoma. MATERIAL: A total of 10 patients (8 males and 2 females) aged from 32 to 74 years (the mean age of 53.3) were included in the study. The enrolment criteria were the following: patients operated for MM or solitary spinal plasmocytoma during the past 7-year period, with the minimum follow up period of 6 months. The procedures were indicated for progressing neurological deficit (Frankel score) and for axial spinal pain (VAS classification), not responding to conservative therapy. The extent of the disease was assessed based on plain x-ray, MRI and whole- body 18F-FDG PET/CT. Paliative vertebroplasty was indicated in patients with no neurological deficit to control pain, paliative laminectomy without stabilization in subjects with partial neurological lesions, with transpedicular fixation in concomitant pathological fractures or kyphotizations. More radical approach, i.e. the procedure included somatectomy, was indicated in patients with solitary plasmocytoma and in procedures on cervical or thoracolumbar regions. Control clinical and MRI examinations were performed at 6 weeks, at 6 months and then at yearly intervals. At the end of the study, the authors evaluated effectivity of the employed surgical procedures, based on all control findings, and the data were compared with prognostic scoring systems in surgery for spinal metastases (Tomita score, Tokuhashi modified score and Bauer score). RESULTS: No local relapses of the tumor or stabilization failure were detected. The effect of surgery on pain control and on prevention of neurological dysfunction was maintained over the follow up period. The authors concluded that all surgical procedures and their radicality were adequate in all subjects. The agreement between the authors approach (the procedure's radicality) and the Tomita score, the Tokuhashi modified score and the Bauer score were recorded in 50% of patients, 80% of patients and in 50% of patients, respectively. DISCUSSION: MM is characterized by increased oseteolysis, which is not followed by new bone formation. Despite successful conservative therapy of MM, the bone defects fail to heal, cause spinal pain and may result in spinal instability. These specific MM signs represent the principal factor in the decision- making process concerning indication for surgery. Furthermore, favourable prognosis, with survival times usually exceeding the required expected minimum survival time of 3-6 months, is yet another reason for indication for surgical therapy in patients with spinal MM. Due to advances in chemotherapy and the use of autologic grafts of peripheral stem cells and radiotherapy, the prognosis of patients have significantly improved in last 10 years. The mean survival time has increased from 2.5 years to 4.5 years. CONCLUSION: 1. Prevention or improvement of neurological dysfuction and pain control are the main indication criteria for surgery in MM. 2. Surgery should be considered in MM with osteolytic spinal disorder and because of favourable prognosis of the disease when surgery is used. 3. Surgical procedures, including paliative methods resulted in sufficient control of spinal stability in all the study subjects. 4. Using all scoring systems for spinal metastases could result in indications for unnecessary more radical procedures. However, Tokuhashi score appeared to be the most suitable existing prognostic scoring system.

[Contribution of whole-body magnetic resonance in the diagnostics of monoclonal gammopathy of undetermined significance, multiple myeloma, and the assessment of Durie-Salmon Plus staging system]. / Prínos celotelové magnetické rezonance v diagnostice monoklonální gamapatie nejistého výnamu, mnohocetného myelomu a stanovení stadia pokrocilosti nemoci s pomocí systému Durie-Salmon Plus.

BACKGROUND: The aim of the study was to assess the contribution of the whole body MRI (WB-MRI) in the diagnostics of monoclonal gammopathy of undetermined significance (MGUS) and initial, asymptomatic form of multiple myeloma (MM), as well as the evaluation of practical usefulness of the Durie-Salmon Plus staging system (D-S Plus). MATERIALS AND METHODS: The analyzed 86-patient cohort consisted of 28 patients with MGUS and 54 patients with newly diagnosed multiple myeloma and 4 patients with solitary plasmocytoma (SP). WB-MRI was evaluated using Magnetom Avanto 1.5 T with the use of virtual whole body coil with sequential acquisition on 7 levels and 2 sequentions--T2 STIR and T1. Based on the number of lesions and the degree of diffuse involvement we assessed the D-S Plus stage, and compared it to the results of standard staging systems according to Durie Salmon (D-S) and International Staging System (ISS). Statistical estimation was done using the Cohen kappa test and McNemara-Bowker test at p < 0.05. RESULTS: In the group of 28 individuals with MGUS, there were 17 (61%) patients fulfilling the IMWG criteria and/orWB-MRI criteria of incipient MM. In 4/17 (23%) patients we described a more advanced stage when comparing D-S Plus to D-S. Nine out of fourteen (64%) patients with MGUS transforming into MM with negative radiological assessment had positive findings on WB-MRI. The character of WB-MRI findings lead in 9/17 (53%) of the patients to the initiation of induction treatment. Stratification according to D-S Plus divided the 54 newly diagnosed patients with MM into stage 1 (16.7%), stage 2 (33.3%) and stage 3 (50%). In 22% there was a shift into a higher stage using DS-Plus in comparison with D-S, in 9% of the patients the shift lead to downstaging. When comparing the results of ISS vs D-S Plus we found that the system based on WB-MRI showed in 41% of the patients higher stage and only in 9% of the patients lower stage. In 13% of MM patients we described extramedulary masses of the tumor, especially in paraspinal region. In 1 of the 4 SP patients the WB-MRI changed the diagnosis into multifocal plasmocytoma. CONCLUSION: WB-MRI is a very contributive imaging method with substantially higher resolution than conventional radiography. It is able to evaluate the grade and the extent of myeloma bone disease. It improves the diagnostic approach in the differentiation of stable MGUS from the phase of malignant transformation into MM. The D-S Plus system proved to be contributive and is competent to become a routine part of diagnostic and stratification algorithms in MGUS and MM.

[18F-FDG PET/CT and 99mTc-MIBI scintigraphy in evaluation of patients with multiple myeloma and monoclonal gammopathy of unknown significance: comparison of methods]. / 8F-FDG PET/CT v diagnostice mnohocetného myelomu a monoklonální gamapatie nejistého významu: srovnání s 99mTc-MIBI scintigrafií.

BACKGROUNDS: Newer imaging modalities, such as 18F-FDG PET/CT and 99mTc-MIBI scintigraphy, have been recently introduced to assess the activity and extent of disease in patients with multiple myeloma (MM) and gammopathy of undetermined significance (MGUS). The aim of our study was to compare the impact of these imaging modalities in the evaluation of MM and MGUS patients. MATERIALS AND METHODS: A total of 101 patients with MM (81 patients) and MGUS (20 patients) were enrolled in the study (21 newly diagnosed and 44 relapsed patients with symptomatic MM, 16 with asymptomatic MM and 20 with MGUS). All patients were without therapy and underwent 18F-FDG PET/CT and 99mTc-MIBI scintigraphy within a maximum interval of 14 days. The scans were classified as normal (N), diffuse (D), and focal or combined (F-FD) pattern. RESULTS: There was no significant difference in the detection of newly diagnosed MM and relapsed patients between the compared methods. 18F-FDG PET/CT performed better than 99mTc-MIBI scintigraphy in the detection of focal lesions (p < 0.039), whereas 99mTc-MIBI scintigraphy was superior in the visualization of diffuse disease (p = 0.042). 18F-FDG PET/CT visualised significantly more focal lesions than 99mTc-MIBI scintigraphy (p = 0.002), both generally in the cohort and when comparing the number of focal lesions per patient. Both the imaging modalities singly or in combination influenced the subsequent clinical management in 17% of patients. In our study, 18F-FDG PET/CT predicted asymptomatic MM and MGUS transformation into more aggressive forms with the necessity to start therapy more often than 99mTc-MIBI scintigraphy. CONCLUSION: 18F-FDG PET/CT appeared to be a better imaging technique than 99mTc-MIBI scintigraphy in the detection of focal lesions in patients with symptomatic MM. 99mTc-MIBI was superior in the visualization of diffuse disease. On the other hand, despite its limited capacity in detecting focal lesions, 99mTc-MIBI scintigraphy still remains the most rapid and inexpensive technique for whole-body evaluation and may be an alternative option when a PET/CT facility is not available.

[Contribution to the evaluation of serum levels of selected biological parameters in monoclonal gammopathy of undetermined significance and in individual clinical stages of multiple myeloma]. / Príspevek k hodnocení sérových hladin vybraných biologických pusobku u monoklonální gamapatie nejistého výnamu a v jednotlivých klinických stadiích mnohocetného myelomu.

INTRODUCTION: This study aimed to measure serum levels of 12 selected parameters in patients with monoclonal gammopathy of undetermined significance (MGUS) and initial, asymptomatic phase of multiple myeloma (MM) to assess their potential benefit in differentiating both conditions. PATIENT SAMPLE AND METHODOLOGY: The analysed sample of 131 patients consisted of 62 patients with MGUS and 69 patients with MM fulfilling the criteria of the International Myeloma Working Group (IMWG). The following techniques were used to assess serum levels: quantitative immunoradiometry (bone-type alkaline phosphatase - bALP and insulin-like growth factor-1 - IGF-1), chemiluminescent enzyme immunochemical reaction (parathormone--PTH), quantitative sandwich enzyme immunoassay (osteopontin--OPN, endostatin (ES), macrophage inflammatory protein-1alpha/beta--MIP-1alpha/beta, angiogenin--ANG a IL-17). Pearson chi2 test and Mann-Whitney U-test were applied during the statistical analysis. RESULTS: The analysis showed statistically significant differences in serum concentrations between MGUS and symptomatic form of MM (Durie-Salmon (D-S) stage 2-3) for: albumin, beta2-microglobulin (beta2-M) and OPN (p = 0.0001 and < 0.0001); osteocalcin for stage 2 (p = 0.006) and MIP-1alpha for stage 3 patients (p = 0.0002). Using the International Staging System (ISS), statistically significant differences between MGUS and all stages of MM (1-3) were identified for albumin and OPN (p = 0.003 and 0.001 and 0.00009, respectively), stages 2 and 3 for beta2-M and ES (p = 0.015 and 0.0001, respectively), stage 2 for ANG (p = 0.014) and stage 3 for MIP-1alpha (p = 0.00001). Statistically significant differences between MGUS and initial, asymptomatic phase of MM (D-S stage 1) was found only for bALP (p = 0.01) and for albumin (p = 0.004) and OPN (p = 0.003) when ISS was applied. Renal function impairment (D-S substage B) showed in comparison to MGUS significant elevation of serum levels of beta2-M (p < 0.0001), OC (p = 0.011), IGF-1 (p = 0.014), OPN (p = 0.003), ES (p = 0.0001), MIP-1alpha (p = 0.0004) and ANG (p = 0.005) and for albumin (p < 0.0001), beta2-M (p < 0.0001) and OPN (p < 0.0001) only when compared to substage A. CONCLUSION: Only albumin and OPN showed useful even though non-specific potential to differentiate MGUS from all ISS stages of MM, beta2-M and ES for ISS stages 2 and 3, the other parameters differed for stage 2 (ANG), stage 3 (MIP-1alpha) only and stage 1 (bALP), stage 2 (OC) and stage 3 (MIP-1alpha) when D-S stratification was applied.

[Analysis of serum levels of selected biological parameters in monoclonal gammopathy of undetermined significance and multiple myeloma]. / Analýza sérových hladin vybraných biologických ukazatelu u monoklonální gamapatie nejistého významu a mnohocetného myelomu.

BACKGROUNDS: The aim of the study was to evaluate the serum levels of 18 selected parameters in monoclonal gammopathy of undetermined significance, and the initial, asymptomatic phase of multiple myeloma, also from the point of view of the potential contribution to the differentiation of these two units. MATERIALS AND METHODS: The analyzed 119-patient group consisted of 59 individuals with monoclonal gammopathy of undetermined significance and 60 patients with multiple myeloma assessed at the time of diagnosis before the start of the treatment. For the evaluation of serum levels we used radioenzyme assay (thymidine kinase), immunoradiometry (IGF-1), enzyme immunoassay (osteocalcin, osteoprotegerin, ICTP), electrochemiluminiscence (PINP), quantitative sandwich enzyme immunoassay (MIP-1 alpha and MIP-1 beta, IL-17, osteopontin, HGF, VEGF, angiogenin, endostatin, syndecan-1/CD138), and for the assessment of serum levels of free light chains kappa and lambda, the Freelite system. Statistical evaluation was done using the Pearson chi-quadrat test and the U-test according to Mann-Whitney (p < 0.05). RESULTS: Statistically significant differences between monoclonal gammopathy of undetermined significance and multiple myeloma were found in the case of serum levels of thymidine kinase (0.0002), ICTP (0.001), MIP-1 alpha (0.002), osteopontin (<0.0001), HGF (< 0.0001), syndecan-1 (<0.0001), and the kappa/lambda ratio (0.0002), while lower significance was found in the case of angiogenin (0.031) and endostatin (0.011). Statistically non-significant differences between multiple myeloma and monoclonal gammopathy of undetermined significance were within the serum levels of IGF-1, osteocalcin, bALP, PINP, OPG, MIP-1 beta, IL-17, parathormon and VEGF. CONCLUSION: Statistical analysis revealed significant differences between monoclonal gammopathy of undetermined significance and multiple myeloma in 9 of the 18 evaluated parameters. However, due to the significant overlapping of the measured values, none of the parameters is unambiguously able to distinguish between the units. A certain contribution in the discrimination of multiple myeloma from monoclonal gammopathy of undetermined significance was found in markedly increased serum levels of thymidine kinase, MIP-1 alpha, osteopontin, HGF and significant pathology of the kappa/lambda index.

The importance of serum levels of selected biological parameters in the diagnosis, staging and prognosis of multiple myeloma.

The study aimed at evaluating the relation of 7 parameters associated with the internal biological properties of myeloma cells and the bone marrow microenvironment to multiple myeloma (MM) stages, distinguishing its initial/asymptomatic phase from monoclonal gammopathy of undetermined significance (MGUS) and assessing their relation to myeloma prognosis. In the studied group comprising 286 individuals (89 MGUS and 179 MM patients), statistically significant differences (Mann-Whitney test) between MGUS and MM at the time of diagnosis were found in the serum levels of HGF (hepatocyte growth factor), VEGF (vascular endothelial growth factor), ICTP (intercellular - carboxy-terminal telopeptide of type I collagen), PINP (procollagen type I N-terminal propeptide), OPG (osteoprotegerin) and syndecan-1/CD138, but not in Fas. Multivariate analysis (logistic regression) revealed an unsatisfactory potential of all the 7 studied indicators to discriminate between MGUS and MM. A deeper analysis showed statistically significant differences between MGUS and the initial/asymptomatic phase of MM (stage 1 according to the International Staging System) only in the cases of syndecan-1 (p=0.001) and Fas (p=0.008). The assessment of initial values of HGF, VEGF, ICTP, PINP, OPG, syndecan-1 and Fas showed a statistically significant relation (log rank test) to the overall survival (OS) in a group of 132 patients treated with conventional chemotherapy only in the cases of syndecan-1 (p=0.0002) and Fas (p=0.018), but in none of the investigated parameters in a group of 74 patients treated with HDT/ASCT (high-dose therapy/autologous stem cell transplantation). The analysis showed that, despite significant differences in serum levels of 6 of the 7 studied parameters found between MGUS and MM, none of the markers may be included in the spectrum of indicators used to distinguish the two conditions. Despite the positive relation, especially of syndecan-1 and, to a lesser extent, of Fas to the OS in patients treated with conventional chemotherapy, these prognostic factors are not applicable to HDT/ASCT.

Thalidomide and bortezomib overcome the prognostic significance of proliferative index in multiple myeloma.

UNLABELLED: We analyzed proliferative index of myeloma plasmocytes (PC-PI) in acohort of 217 patients with multiple myeloma (MM) treated with conventional chemotherapy and biological agents, thalidomide and bortezomib. In the whole group was adifference between overall survival (OS) favoring patients with PC-PI ven after 40 months (median overall survival 25 vs 10months, p= 0.015), whereas in the group treated with thalidomide and bortezomib was no difference, with medians over 39 months. Even patients with low PC-PI profited from the treatment with novel drugs. Presented results suggest that the treatment of MM with novel agents overcomes the prognostic significance of PC-PI and should be used in all MM patients. KEYWORDS: myeloma -prognostication -proliferative index -biological therapy.

[The comparison of serum levels of selected biomarkers in monoclonal gammopathy of undetermined significance and multiple myeloma]. / Srovnání sérových hladin vybraných biologických pusobku u monoklonální gamapatie nejistého významu a mnohocetného myelomu.

BACKGROUND: The aim of the study was the evaluation of serum levels of 12 selected biomarkers in monoclonal gammopathy of undetermined significance (MGUS) and in initial, asyptomatic phase of multiple myeloma, especially from the view of potential differentiation of these conditions in clinical practice. METHODS AND RESULTS: Analyzed group of 268 individuals consisted of 89 individuals with MGUS and 179 patients with MM examined in time of diagnosis before treatment initiation. For evaluation of serum levels were used following methods: radioenzymatic method (thymidinekinase), enzymatic immunoassay (beta2-M, IL-6R, ICAM-1, VCAM-1, ICTP, PINP and OPG) and quantitative sandwich enzymatic immunoassay (HGF, VEGF, syndecan-1/CD138 and Fas). Pearson's chi2-test and test according to Mann-Whitney were used for statistical evaluation (p < 0.05). Wide statistic differences in serum levels of analyzed markers in MGUS vs. MM were detected in case of beta2-M, TK, ICTP, OPG, HGF and syndecan-1 (p < 0.0001), lower differences in case of VCAM-1, PINP and VEGF (p = 0.003, 0.001 and 0.04), and without difference in case of Fas. Except for thymidinekinase (p = 0.014) and syndecan-1 (p = 0.001) was not detected statistically important contrast of measured values in MGUS individuals and in patients with initial, asymptomatic phase of MM (stage 1), but these markers cannot be used in clinical practice due to significant overlap of serum values. Continuous downgrade of serum values of VEGF due to degree of MM progression (stages 1-3 according to Durie-Salmon) was unexpected. RESULTS: Although the significant differences in 9 of 12 evaluated serum levels of selected biomarkers in groups of MGUS and MM were seen, the results revealed that these markers are unprofitable to bring discriminatory potential capable of being used to distinguish between MGUS and initial, asymptomatic phase of MM.

Low-dose thalidomide regimens in therapy of relapsed or refractory multiple myeloma.

Thalidomide has been estimated as a useful drug in therapy of refractory or relapsed multiple myeloma. Recently, several studies have shown very good results in therapy combination of thalidomide, cyclophosphamide and dexamethasone, but still high doses of thalidomide associated with serious adverse events have been used. In our study, we performed low-dose thalidomide regimens; the aim of this study was to verify the effect and to assess their toxicity. For younger patients up to 65 years we used a "CTD-junior" regimen, consisting of oral thalidomide 200 mg daily, pulsed intravenous cyclophosphamide 800 mg on day 1 and pulsed oral dexamethasone 40 mg on days 1-4 and 12-15, for every three weeks. For patients over 65 years, the "CTDsenior" regimen was used, with oral thalidomide 50-100 mg daily (according to tolerability), oral cyclophosphamide 50 mg daily and pulsed dexamethasone 20 mg on days 1-4 and 15-18, for every four weeks. From the group of 97 patients with progressive form of multiple myeloma or with resistance to conventional chemotherapy, 85 patients were evaluated. According to the EBMT criteria, we observed in 8% complete remission (CR), in 50% partial response (PR) and in 22% minimal response (MR). Ten patients (12%) were stabilized and seven patients (8%) progressed. Toxicity of both regimens was mild and well manageable, when weakness, obstipation, neuropathy of lower extremities, glycoregulation worsening and mild leucopenia occurred most often. These results showed that low doses of thalidomide are still effective, when combined with other drugs. Both CTD regimens are safe also for patients with advanced and heavily pretreated multiple myeloma.

[Assessment of plasmatic cell proliferation and apoptosis indexes in the course of multiple myeloma]. / Hodnocení proliferacního a apoptotického indexu plazmatických bunek v prubehu mnohocetného myelomu.

INTRODUCTION: The proliferation and apoptosis indexes are a significant prognostic factor in multiple myeloma (MM). The objective of the study was to assess the indexes in the course of MM from the point of view of the disease activity and potential clinical use. METHOD: A sample of 120 patients was studies, of which 49 at diagnosis, 36 in progression and 35 in remission. The proliferation and apoptosis indexes were assessed according to the achieved treatment response and were compared with subsequent progression in patients in remission. The proliferation potential was measured with the propidium-iodide index (PC-PI/CD138), apoptosis was measured with annexine-V (PC-AI/CD138), assessed by the method of flow cytometry. RESULTS: The sample of49 newly diagnosed patients recorded a decrease in PC-PI (M 2.9-2.1, p < 0.0001) and an increase in PC-AI (M 4.7-8.6, p < 0.0001) at remission. Patients without treatment response showed no difference in PC-PI (p = NS), while PC-AI decreased (M 6.2-3.9, p = 0.004). The sample of36 patients in MM progression with treatment response recorded a decrease in PC-PI (M 2.9-2.2, p < 0.0001) and an increase in PC-AI (M 4.6-8.8, p < 0.019). There was no difference in any of the indicators (p = NS) in cases with zero treatment response. 35 patients in remission with developing progression recorded an increase in PC-PI (M 2.3-2.7, p < 0.0001) and a decrease in PC-AI (M 8.3-4.2, p < 0.0001). CONCLUSION: The study has shown that monitoring PC-PI and PC-AI in the course of different phases of MM is beneficial from the point of view of the assessment of changes in the proliferation activity and the degree of apoptosis of myeloma cells. Monitoring the two parametres allows for timely prediction of disease progression.

International prognostic index (IPI)--a critical comparison with five multiple myeloma staging systems in the group of 270 patients treated by conventional chemotherapy.

In the group of 270 patients with multiple myeloma (MM) treated during 1991-2004 by conventional chemotherapy, the prognostic value and practical utility of IPI (International Prognostic Index) was assessed and compared with five other actual staging systems. Prognostic significance was assessed using the curves of overall survival (OS) according to Kaplan-Meier and log rank test (p<0.05). Good practical utility and prognostic significance of Durie-Salmon (D-S) system was confirmed (p<0.001). Good overall prognostic significance was observed in simple staging systems based on the measurement of beta2-microglobulin and albumin serum levels according to Bataille (p<0.001), SWOG (South West Oncology Group, p<0.001) and IPI (p<0.001). Regardless of a short 5-year duration of the study, the scoring system according to San Miguel enclosing apart from other parameters also propidium iodide proliferation index (PC-PI) of myeloma plasmocytes seems to be promising with very different characteristics of curves of overall survival (p<0.001). Very good prognostic value and easy practical utility were examined in Olomouc staging system (OSS) based on the measurement of beta2-microglobulin and thymidinekinase serum levels (p<0.001). With regard to detection of patients of stage 1, i. e. "low risk", not requiring an immediate initiation of conventional chemotherapy ("wait and see" approach), the most suitable was the system according to D-S, SWOG and IPI (median OS 77, 76 and 77 months). To select a cohort of "high risk" patients, i.e. stage 3, with very unfavourable disease prognosis, the most advantageous was the system OSS and San Miguel (median OS was 5 and 6 months) and/or SWOG system selecting patients of stage 4, i.e. "worst prognosis", with median OS 8 months. It was found that IPI did not meet expectations for effective identification of "high risk" patients (median OS of stage 3 was 20 months) nor for the distinction of different prognosis of patients during initial 25 months of MM course at stage 2 vs. 3. The study indicates that under conditions of common clinical practice and conventional chemotherapy, the staging system according to D-S is still useful, while practical application of SWOG and IPI as simpler alternative to the assessment of clinical stage should be verified by further comparative studies. In harmony with the progress in cytogenetics and molecular biology as well as a prospective requirement of individual target therapy, a future suitable stratification system should be based on parameters of internal biological properties of myeloma tissue and microenvironment of bone marrow, allowing in addition a continuous evaluation of the disease course and the effect of therapy.

Prognostic significance of plasma cell propidium iodide and annexin-V indices and their mutual ratio in multiple myeloma.

The aim of this study was a contemporaneous measurement of plasma cells proliferative and apoptotic activity in patients examined at the time of multiple myeloma (MM) diagnosis before initiation of chemotherapy, focussed on the following aspects: determination of prognostic significance of plasma cell propidium iodide (PC-PI) and annexin-V FITC (PC-AI) indices; optimal cut off of PC-PI and PC-AI with regard to overall survival; calculation of summary kinetic index of plasma cells (PC-PI/AI ratio) for evaluation of its prognostic importance; determination of an index (out of PC-PI, PC-AI and PC-PI/AI) showing the closest relation to prognosis of multiple myeloma. The analyzed 122 patients fulfilling SWOG multiple myeloma criteria were treated by conventional chemotherapy. Plasma cell proliferative activity was measured by means of PC-PI examined by flow cytometry using a DNA/CD138 double staining technique. For detection of plasma cells entering apoptosis (PC-AI), flow-cytometry method with annexin-V FITC and MoAb CD138 was used. The PC-PI median in 122 patients was 2.6(0.4-4.8)%. The sequence prognostic analysis showed that the optimal PC-PI cut off was 2.9% and displayed a significant relationship with overall survival (OS) (p=0.031). The group of 94 patients had PC-AI median of 5.0(1.4-24.5)%. The best statistical significance of the rate of apoptosis related to overall survival was found at cut off value of 4.4% (p=0.022). The median of overall kinetic index of plasma cells (PC-PI/AI) examined in 94 MM patients was 0.5(0.05-2.60) and the overall kinetic index was found to display a very good relationship to OS at the cut off value of 0.71 (p=0.032). All the three indices expressing various aspects of kinetics of plasma cells allow the stratification of patients into two prognostically different groups with statistically significantly different medians of overall survival: good risk - OS still undeterminable at the time of analysis; bad risk - M: OS was for PC-PI 17 months, for PC-AI 23 months and for PC-PI/AI 16 months. The ratio of both indices, i.e. PC- PI/AI, however did not bring any further contribution to overall survival/prognosis evaluation, when compared with single PC-PI and PC-AI. Results of present study indicates that the evaluation of both proliferation and apoptotic activities of plasma cells is important for prognosis thus extending possibilities of initial stratification of MMpatients into groups with different prognostic risk.

[Comparison of serum levels of selected biological parameters in monoclonal gammopathy of undetermined significance and multiple myeloma]. / Srovnání sérových hladin vybraných biologických ukazatelu u monoklonální gamapatie nejistého významu a mnohocetného myelomu.

PURPOSE OF STUDY: The study focuses on evaluation of differences in serum levels of selected biological indicators in monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM), primarily from the viewpoint of potential asset to clinical practice. PATIENTS AND METHODOLOGY: The analyse set of 96 patients consisted of 30 individuals with MGUS and 66 patients with MM examined in the context of the disease diagnosis before therapy commencement. Serum levels of the assessed parameters were examined with the help of radio-enzymatic method (thymidinkinase), radioimmunoanalysis (beta2-micro-globulin, ICTP, PINP), enzymoimmunoessay method (sIL-6R, sVCAM-1, sICAM-1, sOPG and sRANKL) and the technique of quantitative sandwich enzymatic immunoessay (sHGF, sVEGF, bFGF, syndecan-1/CD138 and sFas). Statistical examination was performed by Pearson and Fischer test, chi2-test, or nonparametric U-test pursuant to Mann-Whitney (p < 0.05). RESULTS: Statistically significant differences were found between MGUS and MM in the case of serum level comparisons of sIL-6R (p = 0.02), ICTP (p = 0.001), sHGF (p = 0.001) and syndecan-1/sCD138 (p = 0.001), while no statistically significant differences were present in the case of sVCAM-1, sICAM-1, PINP, sOPG, sVEGF and sFas. During the analysis of frequency of occurrence of abnormal values in the MM and MGUS groups, significant differences were found not only in the case of standard parameters, such as beta2-microglobulin, thymidinkinase, creatinin and albumin, but also in the case of sIL-6R, ICTP, sHGF and syndecan-1, but not in comparisons of the levels of sVCAM-1, sICAM-1, PINP, sOPG, sVEGF and sFas. Attempts at examination of serum levels of sRANKL and soluble form of bFGF failed as too low values were measured. CONCLUSION: The analysis of behaviour of the 10 parameters, mostly intimately related to biological properties of clonal plasmatic cells or to changes of microclimate of bone marrow, effectively contributed to distinction between MGUS and MM only in the case of serum levels of sIL-6R, ICTP, sHGF and syndecan-l (sCD138), i.e. indicators with demonstrable relevance for MM prognosis assessment.

[Imaging methods in diagnosis and monitoring of multiple myeloma]. / Zobrazovací metody v diagnostice a sledování mnohocetného myelomu.

Imaging methods (IM) are important for both the diagnosis and monitoring of the treatment of multiple myeloma (MM). The report discusses radiological IM as well as methods of nuclear medicine. Hole-body screening using simple X-ray pictures is still used in all newly diagnosed cases of the disease, though its validity is significantly higher in chronic forms and primarily in the diagnostics of vertebral compressions. Computer tomography (CT) ideally scans the destructive changes on the compact bone, but it is not very good in showing bone marrow. It is however, invaluable in targeted biopsy or vertebroplasty. Magnetic resonance imaging (MRI) currently has a decisive role mainly in early diagnostics, thanks to its ability to show early changes in the bone marrow. Of critical importance is also indication for MRI in the imaging of structures of the spinal canal and in evidencing epidural propagation of tumour mass. A disadvantage of the method is its inability to show the effects of the treatment immediately following its administration. Contraindications of MRI are also addressed. Among the methods of nuclear medicine, the most important are hole-body 99mTc-MIBI scintigraphy and full-body FDG-PET/CT examinations. 99mTc-MIBI is a sensitive indicator of the biological activity of the disease. It shows the damage to the skeleton caused by the tumour before anatomic changes appear. It reliably differentiates MM remissions from relapses and can be used to determine the optimal position for biopsy puncture. The method is good for monitoring the course of the disease and forecasting the results of the treatment. Its disadvantage is its limited resolution capacity, therefore focal lesions smaller than 10 mm usually escape scintigraphic detection. Similarly to 99mTc-MIBI scintigraphy, FDG-PET/CT examination shows tumorous affection of the skeleton before structural changes appear. It is a highly effective method especially in detecting skeletal damage and extramedullar exhibitions of the disease. The sensitivity and specificity of FDG-PET/CT examination is increased by simultaneous CT examination which is made possible by new generation hybrid instruments. The method, together with 99mTc-MIBI scintigraphy, is very important in the detection of hypo/non-secretional forms of MM. It provides "real time" information on the response of the tumour to treatment and reliably detects the relapse and the remission. An overview is given of recommended examination algorithms for acute and chronic forms and for the monitoring of the treatment of MM, as well as of the importance of all IM for clinical practice.

[Contribution to the evaluation of the relationship between angiogenic cytokines, selected biological parameters and prognostic factors in multiple myeloma]. / Príspevek k hodnocení vztahu angiogenních cytokinu a vybraných biologických ukazatelu k prognostickým faktorum mnohc'etného myelomu.

BACKGROUND: Multiple myeloma is an unusually heterogeneous disease with individually different clinical course, sensitivity to therapy and prognosis. The undoubted limitation of contemporary diagnostic stratification systems is the insufficient credit of those parameters that express the intrinsic biological properties of myeloma cells and the microenvironment of the bone marrow. The aim of this study was to evaluate the relationship of 10 biological parameters to 6 fundamental prognostic factors of multiple myeloma. METHODS AND RESULTS: The analysed group consisted of 66 individuals examined at the time of diagnosis before the start of therapy. For the estimation of serum levels of investigated molecules there were used following methods: REA RIA, ELISA and the technique of quantitative sandwich enzyme immunoassay. For the analysis of proliferative and apoptotic properties of myeloma cells we used propidium iodide (PC-PI) and annexin-V (PC-AI) indices. The statistical estimation was carried out with the help of Pearson and Spearman test, eventually with the help of U-test according to Mann-Whitney. Increased incidence of abnormal serum levels of examined parameter was registered in the case of S-beta2-microglobulin (95.5%), S-thymidinekinase (57.5%), S-sVCAM-1 (78.5%), S-ICTP (87.0%), S-solubile osteoprotegerin (sOPG 76.5%), S-sSyndecan-1 (56.5%) and low index of plasma cell apoptosis (PC-AI, 78.0%). The statistical analysis (Pearson's test) revealed the mutual relationship of serum levels of: S-beta2-microglobulin to sVCAM-1, (r = 0.39, p = 0.002), sICAM-1 (r = 0.33, p = 0.011), sOPG (r = 0.53, p = 0.001), sHGF (r = 0.34, p = 0.006), sSyndecan-1 (r = 0.38, p = 0.003) and sFas (r = 0.42, p = 0.001); S-albumin to sVCAM-1 (r = -0.29, p = 0.036), ICTP (r = -0.33, p = 0.016), sOPG (r = -0.63, p = 0.000), sHGF (r = -0.39, p = 0.003), sSyndecan-1 (r = -0.29, p = 0.042); S-thymidinekinase to sSyndecan-1 (r = 0.46, p = 0.000) and sFas (r=0.29, p = 0.019). There was no relationship of PINP and VEGF to any of the evaluated prognostic factors. There was no relationship found between either of the analysed parameters to PC-PI and PC-AI. With the help of U-test there was found the relationship of sIL-6R to S-beta2-microglobulin (p = 0.001), albumin (p = 0.002) and PC-PI (p = 0.046). CONCLUSIONS: The presented study implies that the traditional algorithm of parameters used in clinical practice for individual characteristics of MM should be enriched with sOPG, sHGF, sSyndecan-1 and sFas.