Monoclonal B-cell lymphocytosis in the population of Slovenian region of Lower Carniola.

INTRODUCTION: Monoclonal B-cell lymphocytosis (MBL) is a premalignant asymptomatic condition characterized by the presence of a small population of monoclonal B lymphocytes (less than 5 × 10(9) /L) in the venous blood of otherwise healthy individuals. Incidence of MBL is higher among first-degree relatives of patients with chronic lymphocytic leukemia (CLL) and also increases with age and among men. We compared the incidence of MBL found in the same environment in both an open general population and relatives of CLL patients. METHODS: Samples were analyzed with 5-parameter flow cytometry using following monoclonal antibodies: kappa and lambda clonality, CD5, CD19, CD20. We analyzed 216 samples of patients in the Slovenian area of Lower Carniola aged 23-88 years. Fifty-four patients who came from families where CLL is present were compared by age and gender to 162 healthy individuals from the general population of the same region. RESULTS: We found 10 cases of MBL (4.6%). In 9.4% of younger (≤ 50 years) first-degree relatives of CLL patients with immunophenotypic characteristics of atypical MBL, we observed a prevailingly normal κ to λ ratio. The difference in incidence of MBL cases between relatives and the general population decreases with age. CONCLUSION: MBL in Slovenia can be coincidentally found with a similar frequency to previously studied populations of other parts of Europe. We found, however, a higher incidence of atypical MBL among first-degree relatives.

Cerebral toxoplasmosis - a late complication of allogeneic haematopoietic stem cell transplantation.

Toxoplasma gondii infection reactivation predominantly occurs among patients after allogeneic haematopoietic stem cell transplantation. Mostly, reactivation occurs during first 3 months after transplant, especially when risk factors are present. We report a case of late cerebral toxoplasmosis reactivation, which was probably triggered by a brief course of corticosteroids, administered for chronic graft-versus-host disease (cGVHD). In the presence of risk factors, such as cGVHD, prophylactic treatment for toxoplasmosis should be reinstituted; Trimethoprim-sulfamethoxasole most probably prevented earlier reactivation of toxoplasmosis in our patient.

T and B lymphocyte rosettes in tonsils of atopic and non-atopic children.

This investigation was carried out to determine the immunological function of the human tonsils, especially the role of lymphocytes of atopic and non-atopic children. An E- and EAC-rosette forming technique is introduced, with semiquantitative evaluation of T and B lymphocytes in the peripheral blood of 56 normal subjects, 28 atopic and 5 non-atopic children of both sexes and different ages. The percentage of T and B lymphocytes in removed palatine tonsils in 28 atopic and 5 non-atopic children was calculated. The percentage of rosette-forming cells in E and EAC suspensions appeared to be independent of age and sex. The percentage of T lymphocytes in peripheral blood was significantly higher than in the palatine tonsils, whereas the percentage of B lymphocytes was significantly higher in the tonsils of atopic and non-atopic children than in the peripheral blood. The mean percentage of B lymphocytes in the tonsils of atopic children was significantly higher than in non-atopic children. The percentage of B lymphocytes was found to be significantly higher in hypertrophic than in small tonsils. Also, in the tonsils with adhesions there was a statistically significantly higher percentage of EAC rosettes than in the tonsils free of adhesions. The percentage of T lymphocytes was statistically significantly higher in the small tonsils.

[Pneumometer values in healthy children and adolescents (author's transl)]. / Pneumometerwerte gesunder Kinder und Jugendlicher. Normalwerte für Knaben und Mädchen von 3--17 Jahren.

On the basis of investigations in 764 healthy children aged 3--17 years (398 boys and 366 girls) normal values for the maximal pneumometric expiration were established. There were definite correlations between the pneumometer values and sex, age, height and weight. The correlation coefficients between the pneumometer values and age or height were about + 0,90. The boys have on average higher pneumometer values than the girls, but the differences are not significant.

Immunological function of tonsils in atopic and nonatopic children, with special reference to E- and EAC-binding lymphocytes.

This investigation was carried out to determine the immunological function of the human tonsils, especially the role of lymphocytes of the atopic and nonatopic children. Authors introduced E- and EAC-rosettes forming technique with semi-quantitative evaluation of T and B lymphocytes in peripheral blood of 56 normal subjects, 28 atopic and 5 nonaptoic children of both sexes and different age. The percentage of T and B lymphocytes in removed palatine tonsils in 28 atopic and 5 nonatopic children was calculated. The percentage of rosettes forming cells in E and EAC suspensions appeared to be independent of age and sex. The percentage of T lymphocytes in peripheral blood is significantly higher than in the palatine tonsils whereas the percentage of B lymphocytes is significantly higher in the tonsils of atopic and nonatopic children than in the peripheral blood. The mean percentage of B lymphocytes in the tonsils of atopic children is significantly higher than in nonatopic children. The percentage of B lymphocytes was found to be significantly higher in hypertrophic than in small tonsils. Also, in the tonsils with adhesions there was a statistically significant higher percentage of EAC rosettes than in the tonsils free of adhesions. The percentage of T lymphocytes is statistically significant higher in the small tonsils.

Molecular epidemiology of catheter-related bloodstream infections caused by coagulase-negative staphylococci in haematological patients with neutropenia.

Catheter-related bloodstream infection (CRBSI) caused by coagulase-negative staphylococci (CNS) is common in haematological patients with febrile neutropenia. As the clinical signs of CRBSI are usually scarce and it is difficult to differentiate from blood culture contamination, we tried to confirm CRBSI by molecular typing of CNS isolated from paired blood cultures (one from a peripheral vein and another from the central venous catheter hub). Blood cultures were positive in 59 (36%) out of 163 patients. CNS were isolated in 24 (40%) patients; in 14 from paired blood cultures (28 isolates) and in 10 from a single blood culture. CNS from paired blood cultures were identified as Staphylococcus epidermidis. Antimicrobial susceptibility was determined and bacteria were typed by pulsed-field gel electrophoresis (PFGE) of bacterial genomic DNA. In 13 patients, the antibiotic susceptibility of isolates was identical. The PFGE patterns from paired blood cultures were identical or closely related in 10 patients, thus confirming the presence of CRBSI. In the remaining four patients they were unrelated, and suggested a mixed infection or contamination. Since CNS isolates from three patients had identical PFGE patterns, they were probably nosocomially spread amongst them.

Comparative study of virological infections in asthmatic and nonasthmatic children.

The author shows complex analyses: clinical, laboratory, X-rays, bronchoscopical, bronchographical and measuring lung function tests as well as the serological examinations in blood serum of both groups of asthmatic and nonasthmatic children with virological infection. The calculation of statistically significant differences between the various diagnostical results of both groups has confirmed that in asthmatic children virological infection of the respiratory tract, pathological findings in X-ray and lung function tests, bronchiectasis and secondary bacteriological invasion occurs statistically significantly more often than in nonasthmatic children.

[Immunologic function of the tonsils in relation to immune competent T and B cells in children]. / Immunologische Funktion der Tonsillen in bezug auf immun-kompetente T- und B-Zellen bei Kindern.

The authors introduced a E and EAC rosette-forming cell technique with semiquantitative evaluation of T and B lymphocytes in the peripheral blood of 56 healthy persons of both sexes and of different ages. The percentages of T and B lymphocytes in the peripheral blood and tonsils of 31 children was also calculated. The percentage of rosette forming cells in E and EAC suspensions appeared to be independent of age and sex. The percentage of T lymphocytes in the peripheral blood is significantly higher than in the tonsils, whereas the percentage of B lymphocytes is significantly higher in the tonsils than in the peripheral blood. The percentage of T lymphocytes was found to be significantly higher in hypertrophic as the mean percentage of T lymphocytes in tonsils of 31 children. Also in tonsils with adhesions there was a statistically significant higher percentage of EAC rosettes than in tonsils free of adhesions.

[Correlation between total serum IgE and the remaining allergologic-diagnostic methods in childhood]. / Korrelation zwischen Gesamt-IgE im Serum und übrigen allergologisch-diagnostischen Methoden im Kindesalter.

The authors found in studing of 60 allergic children on Dermatophagoides Pteronyssinus (D.P.) that besides all clinical allergological tests is extraordinary radioimmunological determinations of serum IgE in allergic patients. In small group of healthy children they found the mean value for serum IgE 499 U/ml (+/- SD - 242). In allergic children with bronchial asthma and vasomotoric allergic rhinitis with positive skin reaction +++ on D.P. and with positive provocation inhalation and nasal test found statistically significantly higher values for serum IgE in chidren with above mentioned diagnostic tests. Detail analyses of determination of total serum IgE showed that is necessary to start to determine also specific IgE with Phadebas IgE test for different inhalative allergens. The greatest advantage of determination of serum IgE in vitro is undengereous technique whereas other allergological diagnostical test especially in childhood are dangerous for live.

[Immunological aspects of infantile tonsilloadenoidectomy (T&A) (author's transl)]. / Immunologische Gesichtspunkte zur Kindertonsillektomie.

Authors studying the frequency of tonsillectomy and adenoidectomy (T&A) have found three times more operations in their country than in the U.S.A. The indication for operation is given by the general practitioner and pediatrician without previous sufficiently long treatment of the underlying disease. The cytological analyses of tonsillar tissues showed a highly active transformation and differentiation of immunologically competent cells and the phagocytes. A close negative correlation between the percentage of the immunologically competent cells and a close positive correlation between the percentage of phagocytes and the frequency of tonsillitis was calculated. The volume, consistency and adhesion of tonsils do not block the highly active transformation and differentiation of the lymphocytes in the tonsillar tissue. The percentage of T-lymphocytes was found to be significantly higher in hypertrophic tonsils, representing a mean value of the percentage of T-lymphocytes in the tonsils of 31 children. Furthermore, in the tonsils with adhesions there was a statistically significant higher percentage of B lymphocytes then in the tonsils which were free from adhesions.

Reticulocyte analysis using light microscopy and two different flow cytometric procedures.

The reticulocyte count is a clinically important indirect indicator of erythropoietic activity of the bone marrow. Reticulocyte enumeration by light microscopy is rather inaccurate and has poor reproducibility. Automation of the reticulocyte count by means of flow cytometry has considerably improved the quality of this investigation. In our study, we compared three methods of establishing the blood reticulocyte number: the microscopic brilliant cresyl blue method and two flow cytometric procedures using thiazole orange (TO), namely FACSort (Becton-Dickinson) and EPICS Profile (Coulter). The aims of the study were (1) to select the most suitable TO concentration to be used with the EPICS Profile cytometer, (2) to determine the correlation between the microscopic method and the two flow cytometric procedures, and (3) to appraise the suitability of flow cytometry for reticulocyte analysis in routine clinical work. According to our results, the most appropriate TO concentration for the EPICS Profile counter is 0.1 mg/L. We observed a good correlation between the three methods tested; the correlation coefficients ranged from 0.82 to 0.87. The mean intra-assay coefficients of variation for the microscopic method and the EPICS Profile and FACSort procedures were 27.5%, 8.4% and 6.3%, respectively.

[Clinical and immunological effect of washing powder enzymes on workers and users (author's transl)]. / Klinische und immunologische Wirkung von Enzymatischen Waschmitteln auf die Arbeiter und Benutzer.

For 141 workers in the washing powder industry in Maribor (Group A) and 68 workers of vegetables' shops to determine the risk of sensitization for house-wives handling enzymes detergents (Group B) as a control group the results of diagnostic investigations as well as analyses of protective measures to proteolytic enzymes are contrasted. Within the Group A eight subgroups according to exposition to proteolytic enzymes powder are evaluated. Especially elucited are the results of skin testing (Scratch-tets with Maxatase, Alcalase and culture extract from B. subtilis) and with proteolytic enzymes powder, the lung function tests (vitalogram, spirogram, pneumometer-test diffusion-test and sometimes the blood gas tests), cytological examinations of nasal mucosa and simultaneously taking a blood sample for determination of concentration of total serum IgE and precipitins to enzymes extracts.

Effects of molgramostim, filgrastim and lenograstim in the treatment of myelokathexis.

Myelokathexis is a very rare form of chronic hereditary neutropenia resulting from impaired neutrophil releasing mechanism in the bone marrow. The recombinant human granulocyte-macrophage (molgramostim) and granulocyte (filgrastim, lenograstim) colony stimulating factors release the mature granulocytes from the bone marrow. We describe a 43-year-old woman suffering from myelokathexis, with the absolute neutrophil count ranging between 0.03 and 1.35 x 10(9)/L. In the period before the introduction of cytokines, the patient had more than 80 major infectious episodes. Since 1991, infections in this patient have been treated with cytokines, given in conjunction with antibiotics. Initially, she received molgramostim in a daily dose of 5 microg/kg subcutaneously, which stimulated the release of granulocytes from her bone marrow, thereby allowing successful treatment of infection. After the development of hypersensitivity, molgramostim was substituted with filgrastim. Finally, lenograstim was given a trial. With all three cytokines, the patient's neutrophil count always attained normal values already 4 hours after subcutaneous application of the drug in a dose of 5 microg/kg, the highest neutrophil levels were measured at 24 hours post-injection, and the neutrophil count was again close to the baseline value 72 hours after the treatment. A slight neutropenia was present 48 hours after the application of filgrastim. We believe that all three cytokines are equally effective in increasing the neutrophil count in venous blood of patients with myelokathexis.

Serum thrombopoietin levels in acute myeloid leukaemia.

To improve our understanding of the regulation of circulating platelet counts (PC) by thrombopoietin (TPO), we studied serum TPO levels and PC before and after myelosuppressive chemotherapy in 12 patients with acute myeloid leukaemia (AML). Serum TPO levels were measured by the quantitative sandwich enzyme-linked immunosorbent assay (Quantikine, RD Systems). At the start of the induction chemotherapy, the patients had a median serum TPO level of 199 pg/ml (range 120-2,150 pg/ml), while 10 to 12 days after the end of chemotherapy, their TPO levels were substantially increased, the median value being 1,907 pg/ml (range 1,049-4,194 pg/ml). The correlation between PC and TPO was statistically significant prior to chemotherapy (p < 0.03) and insignificant after chemotherapy. As a result of chemotherapy, the patients developed aplasia; after the administration of platelet transfusions, their median PC increased to 21 x 10(9)/l (range 5-55 x 10(9)/l), while the median TPO value decreased by 300 pg/ml (range 11-1,125 pg/ml). Our results suggest that platelet mass directly regulates serum TPO levels in acute leukaemia patients prior to chemotherapy and after the administration of platelet transfusions. Serum TPO levels may also be influenced by the cytokine response during complicating infections in patients with chemotherapy-induced cytopenia.

Indices of iron status in patients treated by chronic haemodialysis.

Iron deficiency in patients with end stage renal disease (ESRD) treated by haemodialysis (HD) is difficult to diagnose. The reticulocyte hemoglobin content (CHr) and the percentage of hypochromic red cells (%hypo) are sensitive novel assays for the detection of functional iron deficiency in patients treated with erithropoietin (EPO). In our study thirty-nine chronically hemodialyzed patients were evaluated to determine the value of these two parameters in comparison to the conventional biochemical indicators of iron metabolism. There were significant correlations between CHr and transferrin saturation, CHr and weekly dosage of EPO, and also between %hypo and weekly dosage of EPO. Our data represent superior value of %hypo and CHr to the transferrin saturation and ferritin concentration in detecteng of iron deficiency in HD patients.