RESUMEN
Our study concerned the findings that rat and rabbit heart transplants do not survive after six hours. They become dark, hard and fail to contract within 2 min after reperfusion and never regain their function. We tested the supplementation of solutions for heart transplant preservation with tetrahydrobiopterin (H4B) and L-arginine (L-ARG) to maintain the oxidative and reductive domains of the endocardial NO synthase. We decided to study the excised rabbit hearts preserved in Hank's balanced salt solution (HBSS) at 0 degrees C supplemented with different concentrations of H4B (0, 1, 5, 10 or 100 microM). At desired time intervals, successive pieces stored in the above solutions were warmed to rabbit body temperature in 4 ml of HBSS and maximally agonized by direct application of 20 microl of 200 microM bradykinin (or other agonist) onto the exposed endocardium. Nitric oxide bursts were monitored with a porphyrinic NO sensor lying on the exposed endocardium. Our goal was to find the lowest H4B concentration which would maximally agonize NO. and prolong the time of heart preservation to more than 6 hours. Ten microM are a minimum H4B concentration which achieves maximum prolongation of heart preservation time up to 90 hours. This effect was based upon maximal potentiation of NO. release and minimizing of superoxide production.
Asunto(s)
Trasplante de Corazón , Corazón , Soluciones Preservantes de Órganos , Conservación de Tejido , Animales , Soluciones Isotónicas , Donantes de Óxido Nítrico , Conejos , Ratas , Factores de TiempoRESUMEN
Nitric oxide generated from L-arginine is a messenger for cell-to-cell communication. Abnormalities in nitric oxide release have been implicated in diseases ranging from hypertension and atherosclerosis to septic shock and rheumatoid arthritis. We report here the in vivo and in vitro measurements of nitric oxide in the cardiovascular system using a porphyrinic sensor specific for NO. The sensor has a detection limit 10(-9) M, response time of 0.1-10 ms and diameter of 1-20 microns. Protected by an intravenous catheter or Swan-Ganz catheter, the sensor can be implanted into tissues as well as into the blood stream. Nitric oxide concentrations were measured directly in the heart and also in veins and arteries, ranging in diameter from 100 microns to 5 mm. Nitric oxide production was induced by the action of different physical agents (shear stress, stretching) as well as various chemical substances agonists (bradykinin, acetylcholine, ATP).
Asunto(s)
Sistema Cardiovascular/química , Microelectrodos , Óxido Nítrico/análisis , Animales , Calcio/análisis , Células Cultivadas/química , Endotelio Vascular/metabolismo , Arterias Mesentéricas/química , Miocardio/química , Platino (Metal) , Porfirinas/química , Conejos , Ratas , Venas/químicaRESUMEN
The study assessed the diagnostic value of silver staining method and its possible relevance as an alternative to DNA analysis for the study of cellular proliferation in various leukemias (ALL, AML, CML). Silver staining of nucleolar organizer region-related proteins (AgNORs) was applied to peripheral blood and bone marrow cells. The analysis of S-phase cells was carried out using a FACStar flow cytometer. The mean number of AgNOR dots per nucleus and the percentage of S-phase cells varied according to immunophenotype of leukemic cells, depending on the time of initial diagnosis, remission or relapse. Peripheral blood and bone marrow cells of healthy subjects exhibited less AgNOR dots than leukemic cells. The number of AgNORs in bone marrow cells was higher than that of AgNORs in peripheral blood. Significant differences between ALL and AML, as well as AML and CML in AgNOR quantity were observed. Important increase in AgNOR values was evident in relapsed leukemias and in the CML blast crisis. DNA flow cytometry analyses provided results comparable to those of AgNOR enumeration. The correlation between AgNOR dots and proportion of S-phase cells prompted us to consider that AgNOR count reflects cell proliferation capacity of leukemic cells.
Asunto(s)
Leucemia Mieloide , Proteínas de Neoplasias/análisis , Proteínas Nucleares/análisis , Leucemia-Linfoma Linfoblástico de Células Precursoras , Enfermedad Aguda , Adolescente , Adulto , Anciano , Niño , Preescolar , Femenino , Humanos , Lactante , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Leucemia Mieloide/patología , Leucemia de Células T/patología , Masculino , Persona de Mediana Edad , Región Organizadora del Nucléolo , Fenotipo , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Fase S , Tinción con Nitrato de PlataRESUMEN
Leukemic cells from 10 patients with B-chronic lymphocytic leukemia (B-CLL) were isolated and cultured in the presence of 12-0-tetradecanoylphorbol 13-acetate (TPA) at a concentration of 8 x 10(-7) mol for 72 hours. Cells were analyzed before cultivation and after 72 h of cultivation with and without TPA for changes in surface membrane (Sm) and cytoplasmic (cyt) markers expression, presence of receptor for mouse rosette forming cells (MRFC) and some enzyme profiles. All B-CLL cases studied showed typical B-cell phenotype. TPA treatment induced hairy cell leukemia (HCL) characteristics, given by the membrane CD22 and CD25 expression and TRAP positivity in the majority of the cases tested. Cells had hairy cell-like morphology with more intensive cytoplasmic immunoglobulin (CIg) fluorescence staining, absent receptor for MRFC and increased activity of purine nucleosidephosphorylase. In common these changes indicate that TPA can induce hairy cell characteristics on B-CLL cells in vitro suggesting the more mature differentiation stage of HCL compared with CLL. Furthermore, we originally demonstrated that the CD22, present in the cell membrane after TPA, could be detected in the majority of unaffected B-CLL cells in their cytoplasm. From the technical point of view some intracellular CD markers and Igs of B-CLL cells in viable cells in suspension assayed by flow cytometry are described in this study.
Asunto(s)
Antígenos CD/análisis , Leucemia Linfocítica Crónica de Células B/inmunología , Acetato de Tetradecanoilforbol/farmacología , Animales , Antígenos HLA-DR/análisis , Humanos , Inmunofenotipificación , Leucemia Linfocítica Crónica de Células B/enzimología , Ratones , Células Tumorales CultivadasRESUMEN
In a series of 61 children with newly diagnosed acute lymphoblastic leukemia (ALL) the detection of argyrophilic proteins (AgNORs) in relation to enzymatic profile of leukemic blasts was undertaken. The method of silver staining was used to determine the number of AgNORs per nucleus of cells. The activity of 5'nucleotidase, acid phosphatase and beta-glucuronidase was assessed. The AgNOR proteins quantity varied with immunophenotype and cytochemical profile of leukemic cells. The enzyme 5'nucleotidase is known to be the marker enzyme in beta-precursors ALL and acid phosphatase in T-ALL blast cells. Activity of beta-glucuronidase emerged in lymphoblasts of some cases of ALL in close relation to increased number of AgNOR proteins per nucleus of leukemic cells. Our study indicates the possible importance of beta-glucuronidase involvement and increase AgNOR quantity in the proliferative activity of leukemic cells and thus they are of value in monitoring the risk groups of leukemic patients.
Asunto(s)
Proteínas Nucleares/análisis , Región Organizadora del Nucléolo/química , Leucemia-Linfoma Linfoblástico de Células Precursoras/enzimología , Adolescente , Antígenos Nucleares , Niño , Preescolar , Femenino , Humanos , Inmunofenotipificación , Lactante , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/inmunología , Leucemia-Linfoma Linfoblástico de Células Precursoras/mortalidad , PronósticoRESUMEN
Peripheral blood, bone marrow and/or lymph nodes of 77 patients with T- and B-ALLs/lymphomas were characterized for their surface membrane marker profiles using flow cytometry and fluorescence microscopy. Purine metabolism enzyme activities were compared with membrane immunophenotypes. T and B-ALLs/lymphomas subtypes were defined by the expression of surface membrane antigens detected by the monoclonal antibodies. Based on immunophenotyping we found the following characteristic marker profiles: in T-ALL-CD7, CD2, CD1, CD5, CD3, CD4, CD8, CD38, CD71; in T-NHL-CD7,CD2,CD3,CD4,CD5,CD6; in pre-B ALL-CD10, CD19, CD24, HLA-DR, CD34, in B-ALL-CD19, CD20, CD24, HLA-DR, SmIg with kappa or lamda light chains; in B-ALL-weak SmIg, kappa or lambda, CD19, CD20, CD24, CD5, HLA-DR; in B-NHL-CD19, CD20, CD22, CD24, CD5, more intensive SmIg, kappa or lambda. The cells of leukemic cases tended to have more immature phenotypes than those of lymphoma cases. Analysis of purine metabolism enzyme activities showed that there was a correlation between the values of adenosine deaminase (ADA) and purine nucleoside (PNP) and various types of T- and B-ALLs/lymphomas. ADA levels in B-NHL and B-CLL were lower than those in normal cells, while ADA level in T-ALL, T-NHL, pre-B-ALL and B-ALL was higher (the average 185,92,73,63 pkat. 10(-6)cells, respectively). ADA activity was significantly different between lymphocytes of control group and T-ALL(p<0.01), between T-ALL and T-NHL(p<0.05), between T-NHL and B-NHL(p<0.05) and between T-ALL and B-NHL(p<0.05). PNP activities were lower to those in normal cells. ADA/PNP ratio increased mostly in T-ALL, less in T-NHL, pre-B-ALL and B-ALL (10.8 and 5.3 and 2.2, and 2.0 respectively). ADA/PNP ratio was significantly different between T-ALL and pre-B-ALL(p<0.05) and between T-ALL and B-NHL(p<0.05).
Asunto(s)
Adenosina Desaminasa/metabolismo , Antígenos CD/análisis , Leucemia de Células B/inmunología , Leucemia de Células T/inmunología , Linfoma de Células B/inmunología , Linfoma de Células T/inmunología , Purina-Nucleósido Fosforilasa/metabolismo , Adulto , Niño , Humanos , Inmunofenotipificación , Leucemia de Células B/enzimología , Leucemia de Células T/enzimología , Linfoma de Células B/enzimología , Linfoma de Células T/enzimologíaRESUMEN
Peripheral blood or bone marrow of 24 patients with chronic myeloid leukemia (CML) were characterized for their surface membrane marker profiles using flow cytometry and fluorescence microscopy. Purine metabolism enzyme activities were compared with membrane immunophenotype and cytochemical stains. CML subtypes were correlated with the expression of surface membrane antigens detected by the monoclonal antibodies. On the basis of immunophenotyping we found the following characteristic marker profiles: In stable phase of CML (CML-SP)-CD15, CD11b, CDw65, CD13, in accelerated phase of CML (CML-AP)-CD15, CDw65, CD11b, CD13 and CD33, in myeloid blastic phase of CML(CML-BP-M)-CD13, CD33, HLA-DR, CD11b, CD15, CDw65, in myeloid and lymphoid (mixed) blastic phase of CML (CML-BP-M+L)-CD13, CD33, CD34, HLA-DR, CD11b, CD10 and in chronic myelomonocytic leukemia (CMML)-CD14, CDw65, CD11b, CD33 and HLA-DR. Analysis of purine metabolism enzyme activities showed that there was a correlation between the values of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) and various types of CML. ADA levels in CML-SP, CML-AP and CMML were comparable with those in normal cells. In CML-BP-M, which represents proliferation of less mature myeloid cells (similar to less mature AML subtypes), ADA activity increased and PNP activity decreased. ADA activity was significantly different between control group and CML-BP-M (p < 0.01), between CML-SP and CML-BP-M (p < 0.05). The values of PNP activity were the highest in stable phase of CML (125 pkat. 10(-6) cells) and the lowest (23 pkat.10(-6) cells) in CML-BP-M+L. PNP activity in the other groups corresponded to control values. High ADA/PNP ratio was found in CML-BP-M and CML-BP-M+L (0.7 and 2.0, respectively) in comparison to CML-SP (0.2). It follows from our results that ADA/PNP ratio enables to discriminate between stable and blast phases of CML (p < 0.01). The level of the cytochemical enzymes (CHAE, MPO, SBB, ANAE and 5' NT) varied and reflected the degree of cell differentiation and maturation. CHAE and MPO were characteristic enzymes for CML, ANBE for CMML and 5' NT for CML-BP-lymphoid.
Asunto(s)
Antígenos CD/metabolismo , Antígenos de Neoplasias/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/enzimología , Leucemia Mielógena Crónica BCR-ABL Positiva/inmunología , Purinas/metabolismo , Adenosina Desaminasa/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales , Antígenos de Superficie/metabolismo , Membrana Celular/metabolismo , Femenino , Citometría de Flujo , Humanos , Inmunofenotipificación , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Purina-Nucleósido Fosforilasa/metabolismoRESUMEN
To study the minimal residual disease in acute leukemia patients we used some marker combinations related either to the simultaneous surface membrane and cytoplasmic marker expression, or to the expression of atypical marker combinations, that are absent or extremely rare in normal hematopoietic cells. We investigated to which extent flow cytometric analysis of leukemia-associated marker combination may contribute to sensitive follow-up in patients with acute leukemia. For this purpose dilution experiments were performed, in which artificial mixtures of normal peripheral blood lymphocytes and leukemia cells from a patient with leukemia-associated phenotype were prepared and analyzed for double positive cells. Our results showed that the sensitivity of double color immunofluorescence assay was 3 in 10(4) cells. Sequential studies of residual disease were evaluated in five acute leukemia patients with leukemia-associated markers combinations at diagnosis. In three of them morphologic relapse was preceded by the immunologic detection of small amounts of leukemia cells, while in two other cases, in which no double positive cells for leukemia-associated markers were found, patients are still in hematologic remission. This approach to the study of minimal residual disease could be valuable in monitoring the efficiency of chemotherapy, as well as in evaluating the quality control of bone marrow before autografting. Furthermore, flow cytometric approach can efficiently complete other methods, which are used for more exact definition of remission in acute leukemia patients.
Asunto(s)
Antígenos CD/sangre , Antígenos de Neoplasias/sangre , Biomarcadores de Tumor/sangre , Leucemia-Linfoma Linfoblástico de Células Precursoras/inmunología , Adulto , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Inmunofenotipificación , Leucemia Mieloide Aguda/inmunología , Linfocitos/inmunología , Neoplasia Residual , Sensibilidad y EspecificidadRESUMEN
Peripheral blood cells of twenty-six patients with B cell chronic lymphocytic leukemia (B-CLL) were characterized for their surface membrane and cytoplasmic marker profiles using flow cytometry and fluorescence microscopy. According to surface membrane marker analysis three distinct immunophenotypic subgroups of B-CLL were identified: group I (SIg+, MR+, CD5+, B Ag+, T Ag-; 19 cases), group II (SIg+, MR+, CD5+, B Ag+, TAg+; 3 cases), group III (SIg-, MR+, CD5+, B Ag+, T Ag-; 4 cases). Cells from all patients were positive for the CD19 antigen and at least one of other B cell antigens. Cells from all patients expressed also CD5 and HLA-DR antigens and formed mouse rosettes (MR). Great heterogeneity was found in the membrane and cytoplasmic marking by anti-CD22 MoAb. In four of 23 patients tested, CD22 antigen was expressed in the cytoplasm of CLL cells while it was absent on surface membrane of these cells. This finding was discussed from the point of certain cell heterogeneity in the followed B-CLL cases. Cytoplasmic immunoglobulin (CyIg) detection showed to be very important especially in group III of followed B-CLL cases with undetectable surface immunoglobulins (SIg). Cytoplasmic antigens and immunoglobulin determinations are useful in phenotyping every B-CLL patient, as well as in the immunological study of different maturation stages of B lymphocytes.
Asunto(s)
Antígenos de Superficie/análisis , Inmunoglobulinas/análisis , Leucemia Linfocítica Crónica de Células B/inmunología , Anciano , Anciano de 80 o más Años , Citoplasma/inmunología , Femenino , Antígenos HLA-DR/análisis , Humanos , Inmunofenotipificación , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Formación de RosetaRESUMEN
In the absence of truly leukemia-specific antigen, antigen combinations were identified in leukemia cells that are absent or extremely rare among normal hemopoietic cells. Some of the studied combinations related to the simultaneous surface and cytoplasmic marker expression, others, expressed mainly on cell surface membrane, represented atypical or aberrant combinations. Comparing membrane (m) and cytoplasmic (c) antigen expression (followed in 23 acute leukemia cases), we observed that CD3 could be detected in cytoplasm in the majority of T-ALL cells, while was absent on cell surface membrane where simultaneous expression of more immature T cell markers, such as CD7 and CD5, could be detected. Combination of mCD7/cCD3 could be regarded as a suitable marker of individual T-ALL cells. In cases of B-precursors of acute leukemia cells, leukemia-related combination of mCD19/cCD22 was found, which could characterize a single leukemia cell. The cells in one of 11 AML followed cases were positive for CD13 in cytoplasm, but not on cell surface membrane, where CD33 and other myeloid antigens were expressed. The cells in another two AML cases were positive for CD11 in cytoplasm but not on cell surface membrane, where CD13 or CD33 were expressed. Again, marker combinations of mCD33/cCD13 and mCD13 or mCD33/cCD11, respectively, represent a leukemia-related feature, suitable for tracing single leukemia cells in double immunofluorescence. Acute leukemia defined by the coexpression on most blast cells of antigens classically attributed to different lineages (referred as atypical/aberrant marker combinations) remains a rare event. We isolated a series of 27 (12%) such cases of 225 acute leukemia patients whose cells were immunophenotyped at diagnosis. Myeloid markers were present in T-ALL of two cases, T and B markers were coexpressed in 13 cases, markers of B and myeloid lineage were associated in one case, and T cell and myeloid antigens were found in 10 AML cases; in one AML case (M3 according to FAB classification) an aberrant nuclear coexpression of TdT was observed. In one case of the last group an interesting antigen combination of CD4/CD34 present in AML with monocytic differentiation was observed. When 5 patients with leukemia-associated (aberrant) markers were again analyzed at relapse, the relevant antigen combinations were retained in all of them. In summary, 44 of 50 cases (88%) from our acute leukemia series studied for leukemia-associated antigen combination, both with surface membrane and cytoplasmic marker combinations and those with aberrant markers coexpression allow the detection of minimal residual disease.
Asunto(s)
Antígenos CD/análisis , Antígenos de Superficie/análisis , Biomarcadores de Tumor/análisis , Linfoma de Burkitt/diagnóstico , Leucemia-Linfoma de Células T del Adulto/diagnóstico , Adulto , Anticuerpos Monoclonales , Antígenos CD/sangre , Biomarcadores de Tumor/sangre , Linfoma de Burkitt/sangre , Niño , Antígenos HLA-DR/análisis , Humanos , Inmunofenotipificación , Leucemia Mieloide Aguda/sangre , Leucemia Mieloide Aguda/diagnóstico , Leucemia-Linfoma de Células T del Adulto/sangre , RecurrenciaRESUMEN
A total of 34 AML patients with heterogenous age distribution (from 2 years up to 82 years) were observed. Purine metabolism enzyme activities were compared and correlated with membrane immunophenotype. Analysis of bone marrow and peripheral blood samples based on FAB criteria and immunologic phenotyping of acute myeloid leukemia (AML) provided useful--either confirmatory or contradictory-information on the distribution of M1-M6 patients demonstrating a predominance of M1+M2 and M4 groups (44% and 32.4%, respectively). In contrast, it was demonstrated that less frequent subtypes were M3 and M6 (5.9% and 2.9%, respectively). AML subtypes were correlated with expression of surface antigens detected by the following monoclonal antibodies: CD13, CD33, CDw65, CD11b, CD15, CD14, HLA-DR and CD34. On the basis of immunophenotyping we found the following characteristic markers: M1, M2-CD34, HLA-DR, CD13, CD33, CDw65; M3-CD13, CD33, HLA-DR (negative); M4, M5-CDw65, CD14, CD13, CD33 and HLA-DR. CD14 was confirmed to be a typical marker for discriminating myeloid from monocytoid FAB AML subtypes. Analysis of purine metabolism enzyme activities showed that there is a correlation between the values of adenosine deaminase and purine nucleoside phosphorylase and various immunotypes of AML. High ADA/PNP ratio (> 1.0) was found in M1, M2, M3 subtypes. It was due to the increased level of ADA activity (> 100 pkat.10(-6) cells), though these activities overlapped to a certain extent. It was shown that PNP activity simultaneously decreased. With maturation of cells within AML lineage ADA activity decreased and PNP activity increased. This corresponded with ADA/PNP ratio that was < 1.0 in cells of more mature AML subtypes. We found that the enzymatic values were characteristic mainly in cells of M5 (monocytic) AML subtype and were characterized by decreased values of ADA activity with a simultaneous increase in PNP activity. It follows from our results that ADA/PNP ratio enables to discriminate between myeloid and monocytoid subtypes of AML.
Asunto(s)
Adenosina Desaminasa/biosíntesis , Leucemia Mieloide/enzimología , Leucemia Mieloide/inmunología , Purina-Nucleósido Fosforilasa/biosíntesis , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antígenos CD/análisis , Niño , Preescolar , Cromatografía en Papel , Técnica del Anticuerpo Fluorescente , Antígenos HLA-DR/análisis , Humanos , Inmunofenotipificación , Persona de Mediana EdadRESUMEN
Purine metabolism enzyme pattern, cytochemical markers and clinicopathologic features of common acute lymphoblastic leukemia antigen (cALLA; CD10)-positive, CD10-negative T acute lymphoblastic leukemia (ALL), and cALLA-positive non-T, non-B ALL (common ALL; C ALL) of children were compared. The results of immunophenotyping of blast cells in 61 children with ALL who were treated and followed during the last 7 years at the Second Pediatric Clinic in Bratislava are presented. The aim of our study was to determine the correlation of CD10 marker expression with purine enzyme activities and clinical course in ALL of children. Immunologic phenotype performed by a panel of monoclonal antibodies in indirect immunofluorescence assay revealed 3 main ALL groups: Common ALL (C ALL), T ALL and CD10+ T ALL (C + T ALL). An additional exact cytochemical marker analysis was performed in these three ALL immunologic subtypes. Two enzymes of purine metabolism, i.e. adenosine deaminase (ADA) and purine nucleosidephosphorylase (PNP) were investigated in blast cells by paper radiochromatography. Life-table analysis revealed significant prognostic differences with regard to event-free survival and overall survival in followed groups of ALL patients. Our results showed a rather high frequency of mixed (C + T) ALL phenotype. The characteristic T ALL enzyme pattern (high ADA, low PNP) was present not only in T, but also in CD10+ T ALL blast cells. The T cell marker showed to be dominant in the determination of clinical course and prognostic significance in children with ALL; children with T and CD10+ T ALL phenotype, in contrast to C ALL phenotype, experienced more frequent relapses and a shorter event-free survival.
Asunto(s)
Adenosina Desaminasa/análisis , Antígenos de Diferenciación/análisis , Antígenos de Neoplasias/análisis , Leucemia-Linfoma de Células T del Adulto/enzimología , Purina-Nucleósido Fosforilasa/análisis , Fosfatasa Ácida/análisis , Adolescente , Niño , Preescolar , Femenino , Humanos , Lactante , Leucemia-Linfoma de Células T del Adulto/inmunología , Leucemia-Linfoma de Células T del Adulto/mortalidad , Masculino , Neprilisina , Fenotipo , Tasa de SupervivenciaRESUMEN
The chronic lymphocytic leukemia, the prolymphocytic leukemia and the hairy cell leukemia of B-cell immunophenotype are closely related disorders, but differ in their cytomorphological and clinical features. In an attempt to differentiate further among these forms of leukemia some immunological and cytochemical markers were studied. Simultaneously we measured adenosine deaminase and purine nucleosidephosphorylase activities by a method of paper radiochromatography in peripheral blood/bone marrow leukemic cells from 23 patients with chronic lymphocytic leukemia, 5 patients with prolymphocytic leukemia, one with prolymphocytoid transformation of chronic lymphocytic leukemia and 15 patients with hairy cell leukemia. The mosaic of immunological and cytochemical markers based on the sum of positive and negative features allowed for the correct diagnosis in a majority of cases. From the number of 43 investigated cases we found the typical enzyme patterns in 39 of them. On the basis of purine enzyme activity we were able to differentiate between chronic lymphocytic leukemia versus prolymphocytic and hairy cell leukemia. In one patient with chronic lymphocytic leukemia we could detect very early stage of prolymphocytoid transformation by increased activity of purine nucleosidephosphorylase activity. There were only two patients with chronic lymphocytic leukemia who were assigned to the prolymphocytic leukemia on the basis of purine nucleosidephosphorylase activity and two patients with hairy cell leukemia with atypical enzyme pattern attributable to the nonrepresentative number of pathological cells in the peripheral blood. Our study showed that purine nucleosidephosphorylase activity in leukemia cells may be useful as an additional parameter in the distinction of prolymphocytic from lymphocytic leukemia and that it may represent an enzymatic marker for early detection of prolymphocytoid transformation of chronic lymphocytic leukemia. Characteristic purine enzyme pattern was found also for diagnostic confirmation of hairy cell leukemia.
Asunto(s)
Adenosina Desaminasa/análisis , Linfocitos B/inmunología , Biomarcadores de Tumor/análisis , Leucemia de Células Pilosas/enzimología , Leucemia Linfocítica Crónica de Células B/enzimología , Leucemia Prolinfocítica/enzimología , Purina-Nucleósido Fosforilasa/análisis , Linfocitos T/inmunología , Adenosina Desaminasa/sangre , Anticuerpos Monoclonales , Antígenos de Diferenciación/análisis , Biomarcadores de Tumor/sangre , Médula Ósea/enzimología , Técnica del Anticuerpo Fluorescente , Humanos , Cadenas Ligeras de Inmunoglobulina/análisis , Cadenas kappa de Inmunoglobulina/análisis , Cadenas lambda de Inmunoglobulina/análisis , Leucemia de Células Pilosas/sangre , Leucemia de Células Pilosas/inmunología , Leucemia Linfocítica Crónica de Células B/sangre , Leucemia Linfocítica Crónica de Células B/inmunología , Leucemia Prolinfocítica/sangre , Leucemia Prolinfocítica/inmunología , Fenotipo , Purina-Nucleósido Fosforilasa/sangre , Formación de RosetaRESUMEN
Histologic sections of the aortic arches from guinea pigs were stained with nile red to verify the possibility of quantitating the earliest changes in the arterial wall. Nile red fluorescence intensity was quantitated by conversion to electric signal. Fluorescence in a group fed cholesterol and casein was significantly higher than that in a control group. Its dependence on the logarithm of serum cholesterol concentrations between 1.33 and 7.67 mmol/l was found. Small fluorescence intensity differences far beyond the possibilities of visual observation were detected by the method.