Prognostic significance of bone-marrow pattern and immunophenotypic score in B-chronic lymphocytic leukemia at diagnosis.

INTRODUCTION: The clinical course and outcome of B-CLL is various and so far unpredictible. Defining prognostic parameters potentiating division of patients in groups with favorable and unfavorable prognosis which could help the benefit assessment of early treatment, improve treatment effects, and potentiate treatment modification for each patient. AIM: To analyze the bone-marrow (BM) pattern and immunophenotypic score at diagnosis of B-CLL and determine the correlation of BM pattern with the clinical stage of disease and immunophenotypic score. METHODS: A sample of 40 untreated patients with B-CLL was divided into two groups: group with clinical stage Binet A and group with clinical stages Binet B and Binet C. BM patterns were observed as a diffuse, interstitial, nodular and mixed. BM immunophenotyping included CD5, CD23, CD22, and CD20 as an indirect indicator of FMC7. RESULTS: The overall sample mean age was 62.88 years +/- 11.10, without significant difference in the age of two compared groups (63.15 +/- 10.53 years vs. 62.60 +/- 11.50 years) (t = 0.16, df= 38, p = 0.88). Proportion of men was significantly higher in stages Binet B and C (12/20) compared to stage Binet A (5/20) (Z=2.24, p=0.025). The percentage of women was higher than men in Binet A stage (75% vs. 25%). The BM patterns in Binet A stage were observed as follows: mixed 50% (10/20), interstitial 30% (6/20), nodular 15% (3/20) and diffuse 5% (1/20). The BM patterns in Binet B and C stages were observed as follows: diffuse 50% (10/20), mixed 40% (8/20), interstitial 5% (1/20) and nodular 5% (1/20). Clinical stage and the BM patterns were significantly associated (c2=8.02, p=0,005). The chance for non-diffuse patterns was 19 times higher in stage Binet A compared to stages Binet B and C, respectively, analyzing 95% CI at least 2 times higher (95% CI: 2.02-866.6). Immunophenotypic score in total sample was observed as follows: score 4: 5% (2/40), score 3: 72.5% (29/40), score 2: 20% (8/40) and score 1: 2,5% (1/40). Immunophenotypic score 3 and > 3 had 77.5% of patients (31/40), but there was no significant association between the immunophenotypic score and the BM patterns (c2=0.76, p=0.38). CONCLUSIONS: Diffuse BM pattern was significantly associated with the clinical stages Binet B and C, compared to non-diffuse BM patterns which were significantly associated with the clinical stage Binet A. Diffuse BM pattern represent the parameter of progressive disease compared to the non-diffuse BM patterns which are more often represented in stable disease. Immunophenotypic score improves diagnostic accuracy of B-CLL, but should not be used as a prognostic parameter of B -CLL.

Morphometric angiogenesis parameters for indolent and aggressive non-Hodgkin's lymphoma.

There is much evidence about importance of angiogenesis in development and progression of solid tumors. The role of angiogenesis, as an indicator of higher malignant potential in non-Hodgkin's lymphoma, is not clear at the moment. Morphometric characteristics of microvessels in lymph node sections, in previously untreated patients with small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL) and diffuse large B-cell lymphoma (DLBCL), were studied and relationship between angiogenesis and histological malignancy grade of NHL was also evaluated. Lymph node biopsies samples of 30 newly diagnosed patients with SLL/CLL (n=30) and DLBCL (n=30) were studied. All samples were fixed in 10% buffered formalin solution and embedded in paraffin. Microvessels were visualized by immunohistochemical staining for anti F-8 antibody. In the area showing the most intense vascularization (i.e. the "hot spot"), microvessel density (MVD), total vascular area (TVA), as well as the size related parameters were estimated, by using image analysis program "analysSIS'. Number and size-related microvessels angiogenic morphometric parameters were statistically higher in group with DLBCL compared with SLL/CLL: MVD (p = 0.002), TVA (p < 0.0001), area (p < 0.0001), perimeter (p < 0.0001), minor axis length (p < 0.0001) and major axis length (p < 0.0001). It is to be noted that positive correlation existed between TVA and MVD in DLBCL and SLL/CLL. The present study supports the view that angiogenesis correlate with histological grade of NHL.

The role of predictor factors in patients with non Hodgkin lymphoma in relation to the applied therapy.

UNLABELLED: Lymphoma are defined as malignant neoplasms of immune system, or in other words the neoplasms of lymphocytes and their precursory cells which form the basic cellular elements of the immune system. The heterogeneity of lymphoma from the aspect of morphology and the aspect of clinical manifestations demanded the identification of all possible parameters which could influence the course and outcome of the disease on every single level. The aim of our study was to investigate which of the prognostic factors confirms the statistical importance of the remission state and overall survival and if the patients treated with standard PHT (poly chemotherapy) protocols with the addition of anti-CD20 antibody show more clinical benefit. PATIENTS AND METHODS: Considering the applied protocol, patients were divided in two groups: group of patients treated with PHT protocol CHOP (Cyclophosphamide, Adriamycin, Oncovin, Pronison) and the group treated with PHT protocol with the addition of anti-CD20 antibody (Rituximab). RESULTS: There was no statistically significant difference in values of Karnofsky score between patients in CHOP and CHOP+R groups before the therapy (U=133; Z = -1.87; p = 0.06), although this difference was on the border of statistical significance. The values of Karnofsky score before and after the therapy in CHOP group showed no significant difference (Wilcoxon Z = -1.62; p = 0.11), but CHOP+R group pointed significant differences before and after the treatment, (Wilcoxon Z = -2.34; p = 0.02) with generally higher values after the therapy. Looking from the aspect of B-symptomatology in CHOP group, there was a significant reduction of B-symptoms (50%; %95 CI = 19-50; p = 0.002) after the therapy (7/20; 35%) in comparison with patients before the therapy (17/20; 85%). According to those results, the reduction of symptoms in CHOP +R group was more evident (60%; 95% CI = 28-60; p < 0.001) after the therapy (3/20; 15%) comparing to the state before the therapy (15/20; 75%). Comparison of LDH values in the complete sample demonstrated that the value of LDH before the therapy was significantly higher (Z = -3.76; p < 0.001). There was a significant difference in number of patients in relation to the category of IPI prognostic index (chi2 = 10.38; df=3; p < 0.02), pointing out most of the patients with low intermediary risk in CHOP+R group, and largest number of patients with high and high intermediary risk in CHOP group.

Cystatin C in sera of patients with aggressive non-Hodgkin B-cell lymphoma.

AIM: To investigate the cystatin C levels in sera of patients with aggressive non-Hodgkin B-cell lymphoma. METHODS: The levels of cystatin C in sera of lymphoma patients and control group consisted of healthy individuals, were measured by using specific sandwich-type ELISA. For each patient the clinical stage of disease was determined according to Ann Arbor staging system for lymphomas. RESULTS: Our study shows that mean cystatin C serum level in the patients group (1056 +/- 65 ng/mL) was significantly higher when compared with the mean level of the healthy control group (819 +/- 28 ng/mL) (P = 0.001). Mean cystatin C level of the group with clinical stages III and IV (1255 +/- 109 ng/mL) was significantly elevated when compared with the mean level of the group with clinical stages I and II (896 +/- 51 ng/mL) (P = 0.03). CONCLUSION: This finding points out a connection between inhibitor level and aggressive behaviour of lymphoma and could be considered for further strategies of prognosis of the disease.

[Case report of successful treating of refractory form of chronic lymphatic leukemia in Binnet C stage by using anti-CD52 antibodies]. / Uspjesno lijecenje refrakterne forme hronicne limfaticne leukemije u stadiju Binnet C primjenom antiCD52 antitijela.

Chronic lymphatic leukemia (CLL) is the most frequent type of leukemia in western world, and a choice of treatment modality depends on current stage of disease. Clinical condition of patient considered as Binnet C stage, requires treatment. Standard polyhemiotherapy (FC protocol) does not always warrant adequate and satisfactory response. This case report reviews the patient with CLL in Binnet C stage, who did not respond on FC protocol in expected way, meaning, hematological and medullar response was not detected. Twelve weeks therapy of monoclonal antiCD52 antibody (MabCampath) was than applied, resulting in normalization of all parameters of disease activity, which was desired effect of the therapy. Administration of monoclonal antiCD52 antibody is justified in case of resistance on conventional previously applied means of therapy.

Cystatin C as a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.

The concentration of cysteine protease inhibitor cystatin C was determined in sera from 59 patients with non-Hodgkin B-cell lymphoma using ELISA. The sera from 43 age and sex matched healthy blood donors served as controls. Cystatin C was significantly increased in sera of patients without therapy (mean 1136+/-SE 105.7ng/ml, p=0.00001) and with therapy (mean 1073+/-52ng/ml, p=0.001) compared to controls (mean 819+/-28ng/ml). The highest levels were determined in sera of patients with a relapse (mean 1680+/-196ng/ml). By using immunofluorescence staining and confocal microscopy we determined immature dendritic cells as a major population of cystatin C positive cells in affected lymph nodes. Our study reports for the first time that cystatin C is a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.

[Non Hodgkin lymphoma in the North-East Bosnia--changes in biological aggressiveness and primary presentation of the disease]. / Non Hodgkin limfom u Sjeveroistoinoj Bosni--promjene u bioloskoj agresivnosti i nacinu primarne prezentacije.

INTRODUCTION: NHL are the group of lymphoproliferative neoplasms which incidence is in constant increase. The aim of the paper is the analysis of the immunophenotypes, clinical distribution and ways of primary presentation and biological aggressiveness of NHL in the North-East Bosnia. It especially illustrates the post war period from 1998 to 2002. METHODS: Two hundred and twenty-eight patients with pathohistologically proved NHL are analysed in the paper. The tested group consisted of 142 patients with diagnosed NHL in the period between 1998 and 2002, and the control group consisted of 86 patients whose NHL was proved in the period between 1987 and 1991. These factors were analysed: immunophenotype NHL by immunohistochemical method of indirect three-stage immunoperoxidase with streptovidin, the type of NHL, the degree of biological aggressiveness of NHL, the way of primary presentation and the clinical phases of distribution of the disease according to the age, sex, profession and the habitat (urban or rural) of the patients. The prevalence and the incidence of the disease in the region of the North-East Bosnia was determined. The statistical analyses of the date was performed by the average values, standard deviations and chi2 test. RESULTS: The total number of patients with NHL in the North-East Bosnia was quite bigger in the test group than in the control group with the incidence of 2.6/100,000 inhabitants in 1989, and 6.91/100,000 inhabitants in 2002. There were 80 men (56%) and 62 women (44%) in the test group, and 55 men (64%) and 31 women (36%) in the control group. The average age of the patients was 55.78 +/- -15.1 years. Statistically significant decrease of patients was noticed in the groups of industrial and agricultural workers in the test group comparing with the control group (9% against 15% and 4% against 19%, p < 0.005). At the same time the significant increase of the number of patients was found in the test group from the industrial regions of Lukavac (15 against 4 patients, p < 0.005), Tuzla (49 against 19 patients, p < 0.005) and Zivinice (14 against 3 patients, p < 0.005). Diffuse Large Cell Lymphoma (DLCL) was dominant in the test group with total of 73 patients (51%), and Small Cell Lymphoma was dominant in the control group with total of 33 patients (38%). Statistically significant increase of both DLCL and MALT lymphoma is found in the test group (p < 0.025), and the most frequent were patients with IV-B (18%), I-AE (15%) and II-BE (12%) clinical stadium, while in the control group the most frequent number of patients was in the clinical studia III-B (19%), II-B (14%) and IV-B (14%). The most prevalent stadium in both groups was B fenotype with 94% of cases in the test and 93% in the control group. The aggressive lymphomas are predominant in the test group (62%) while the indolent ones are predominant in the control group (64%). In both groups the most of the patients were with nodal primary presentation (51%). In the test group there was a significant increase of aggressive lymphoma in both men and women (p < 0.01). DISCUSSION: In recent years, all NHL studies have shown an increase of the incidence in patients, especially in older age while the increase is not found only in infants. In our study the highest percentage of the incidence of NHL occurs in patients who are 55 and older that exactly matches the literature date. There is an increase of extranodal primary presentation (28%) in test group in comparison with the control group (18%). The distribution of the patients is not significantly different from the findings of the European Oncology Association from 2003 where it says that there are 54% of patients with primary nodal presentation, 34% of them with primary extranodal presentation and 12% combination of the two presentations. Recently, the most frequent presentation worldwide is in stomach, which is also proven in our research. In the test group the number of MALT and DLCL lymphoma located in stomach is in increase. In the last few years the dominant type of NHL worldwide is DLCL. Therefore the significant increase of the patients with this disease is noticed in our test group. The continuous increase of patients with aggressive NHL in the recent years has been noticed especially Lukavac, Tuzla and Zivinice. These are the regions where the electric plant and the chemical industry factories are located. CONCLUSION: The incidence of NHL in the region of the North East Bosnia follows the world trend of the general increase of the NHL incidence including the significant increase in number of aggressive lymphoma. The frequency of DLCL and MALT lymphoma is evidently in increase. The significant changes in primary presentation of the disease have not been noticed. B phenotype of NHL is predominant in both periods of testing.

[Clinical characteristics of pruritus in hemodialysis patients]. / Klinicke karakteristike pruritusa bolesnika na hemodijalizi.

AIM: Aim of the research was to analyze clinical characteristics and most important risk factors of uremic pruritus. PATIENTS AND METHODS: A total of 151 patients on chronic hemodialysis (CHD) during at least 12 months were analyzed. Thorough history was taken for pruritus, its presence and localization, sleep disorder and neuropathic symptoms. On physical examination, attention was focused on the skin. Laboratory tests includes blood cells count, serum urea, creatinine, electrolytes, aminotransferases, alkaline phosphatase and proteins. The dose of dialysis was followed by Kt/V. On statistical analysis, t-test and chi2 test were used. RESULTS: Pruritus was present in 85 (56%) patients, 41 women and 44 men, mean age 53.56+/-13.36 (26-81) years, mean time on CHD 78.36+/-55.02 (12-268) months. There were 66 (44%) patients without pruritus, 32 women and 34 men, mean age 50.35+/-13.76 (22-73) years, on CHD for 58.64+/-50.40 (12-187) months. Although the patients with pruritus were somewhat older and longer on CHD, there was no significant difference either in sex structure or distribution according to primary renal disease. In the group with pruritus there were significantly more anuric patients (43 vs. 22) (p<0.01). The patients with pruritus had a higher rate of sleep disorder (NS), calcium deposits in soft tissues and blood vessels (NS) and clinical neuropathy (p<0.01). Skin changes were found in almost all patients with pruritus (93%), which differed significantly from the patients without pruritus (48%) (p<0.005). The mean value of Kt/V was 1.23+/-0.35 in patients with pruritus, and 1.34+/-0.41 in those without pruritus (NS). Kt/V higher than 1.4 was significantly less frequently recorded in patients with pruritus than in those without pruritus (55%) (p<0.005). The red cell and white cells count, serum hemoglobin, calcium, phosphorus and their products, aminotransferases, bilirubin, alkaline phosphatase and proteins were approximately the same in both groups of patients. CONCLUSION: The loss of residual renal diuresis, Kt/V below 1.4, presence of calcium deposits and neuropathy were the most common risk factors for the extent of uremic pruritus in our CHD patients.