Search PubMed⌕ Search

Biomedical subjects

R Berger

Publications and source records attributed to R Berger.

At least 703 records · Page 39Linked to original sources

Promyelocytic blast crisis of chronic myelocytic leukemia with both t(9;22) and t(15;17) in M3 cells.

A blast crisis with the features of promyelocytic leukemia (M3) occurred during the evolution of chronic myelocytic leukemia (CML) with the t(9;22) translocation. This rare form of transformation was confirmed by means of cytologic and electron microscopic examination. Cytogenetic studies showed two simultaneous translocations t(15;17) and t(9;22) in the promyelocytes. After intensive chemotherapy, a complete remission was obtained and only karyotypes with t(9;22) translocation were present. These data confirm the specificity of the t(15;17) translocation in malignant promyelocytic proliferation and provide evidence for a second genetic event in the genesis of blast crisis occurring in a committed cell belonging to the abnormal population defined by the Ph1 chromosome.

Bone Marrow↗

Dihydropyrimidine dehydrogenase deficiency leading to thymine-uraciluria. An inborn error of pyrimidine metabolism.

Three unrelated patients with excessive thymine-uraciluria due to dihydropyrimidine dehydrogenase deficiency are described. Excretory values (mmol/g creatinine) were: uracil 2.0-10.5, thymine 2.3-7.5, 5-hydroxymethyluracil 0.2-0.9. Orally administered (index patient) uracil and thymine were excreted for the greater part whilst dihydrouracil and S-dihydrothymine were mainly metabolised. Dihydropyrimidine dehydrogenase activities (nmol X h-1 X mg-1 protein) in leucocytes were 0.04, 0.01 and less than 0.01 in the patients, 0.31-1.66 in their parents, and 1.01-4.46 in controls (n = 4). The patients presented with a non-specific clinical picture of cerebral dysfunction.

Adolescent↗

Subacute and chronic myelomonocytic leukemia in children (juvenile CML). Clinical and hematologic observations, and identification of prognostic factors.

The clinical and hematologic characteristics of 38 children with subacute and chronic myelomonocytic leukemia (S & CMMOL) are described, and the prognostic significance of these characteristics as recorded at diagnosis is reported. The common and distinctive feature of these children was the excessive proliferation of cells of neutrophilic and monocytic series. The disease predominated in younger children, 95% were younger than 4 years, and boys were more affected than girls (22/16). The onset of the disease was heralded most often by acute or subacute symptoms. Splenomegaly was the most common physical finding at diagnosis. Leukocytosis was usually under 100 X 10(9)/l. Monocytosis and granulocytosis were often associated with normoblastosis, and, in some cases, with moderate blastosis (less than or equal to 30%). Severe anemia and marked thrombocytopenia were found in about one third of patients, increased fetal hemoglobin levels in 53%, and increased gamma-globulin levels in 50% of cases. The Philadelphia chromosome was absent in all blood and marrow cell karyotypes. Thirty-three of 38 patients were treated with moderate or intensive chemotherapy, and in all cases treatment never resulted in a complete remission. Terminal acute leukemia occurred in 11 cases. Of the 38 patients, 29 have died (median survival time, 16 months). Initial characteristics predicting a short survival (log-rank test) included: older age (greater than or equal to 2 years) (P less than 0.001), hepatomegaly (P less than 0.05), bleeding (P less than 0.001), thrombocytopenia (P less than 0.01), high counts of blasts and normoblasts in peripheral blood (P less than 0.01, P less than 0.01). Sex, infections, cutaneous manifestations, lymphadenopathy, degree of splenomegaly, hemoglobin levels, fetal hemoglobin, leukocyte counts, percent of blasts in bone marrow, and serum gamma-globulin levels were of no prognostic value. When survival was plotted on a semilogarithmic scale, a change in death rate was evident at the second year of survival suggesting that there may be two subgroups of patients with myelomonocytic picture, one with very rapid, and another with a slower rate of mortality. A stepwise discriminant-function analysis was performed in an attempt to distinguish between those children who lived less than or equal to 2 years and those who lived longer. A linear combination of variables which best discriminated between these two subgroups was found. Nearly all patients could be classified as a short-survivor or long-survivor on the basis of age and platelet, blast, and normoblast counts in peripheral blood.(ABSTRACT TRUNCATED AT 400 WORDS)

Bone Marrow Examination↗

Interferon induction in human mixed leukocyte-tumor-cell reactions: evidence for restriction to a certain lineage expressing glycophorin A.

This study reports the induction of interferon (IFN) in human mononuclear cells (MNC) by hematopoietic tumor cells. Only 3 out of 15 cell lines were capable of inducing IFN in the mixed leukocyte/tumor-cell reaction (MLTR). K562, a pluripotent stem cell line and DUTKO-I, a hybrid between K562 and Daudi (Burkitt lymphoma) induced high levels of antiviral activity (ranging from 50-440 units IFN/ml). PUTKO-I, a hybrid between K562 and P3HR-I (Burkitt lymphoma) induced very low levels of IFN (10 units/ml). This antiviral activity was produced by HLA-DR + adherent cells as revealed by different cell separation techniques, and shared well-known properties of IFN gamma (isoelectric point, inactivation by anti-human IFN gamma antibodies; species-restricted protection). MLTR-induced IFN induction could be blocked by enzyme treatment of tumor cells, but was still present when glutaraldehyde-fixed cells were used for induction. Analysis of the cells by flow cytometry for expression of glycophorin A (GpA) revealed that expression of GpA correlated with the ability to induce antiviral activity in MLTR. Furthermore, isolated GpA could be used as a stimulant as well and the response to either K562 cells or soluble GpA was enhanced up to ten-fold by the addition of a GpA-specific monoclonal antibody.

Cell Communication↗

Rapid and early determination of sex using trophoblast biopsy specimens and Y chromosome specific DNA probes.

The feasibility of determining sex by analysing deoxyribonucleic acid (DNA) with two probes specific for Y chromosomes was shown using DNA obtained from samples of blood from 30 non-related males and females of different ethnic origin. The DNA was spotted on nitrocellulose filters and hybridised with both a repetitive (P1) and a unique (49f) sequence specific for the human Y chromosome. A strong positive signal with both probes indicated the presence of male DNA. The sex of 12 fetuses was then similarly determined by molecular characterisation of DNA from trophoblast biopsy specimens. Chorionic samples were obtained in seven cases before termination of pregnancy in the first trimester and the aborted embryos subjected to karyotyping and sex chromatin analysis. In the five other cases samples were obtained from placentas obtained during caesarean section. Results of hybridisation were compared with those from cytogenic studies and actual sex at birth. The sex of all 12 fetuses was determined correctly by hybridisation.

Chorionic Villi↗

A simple device to obtain high local concentrations of material sorted by flow cytometry for biochemical or morphological analysis.

A system is described which allows us to collect flow cytometry sorted objects onto a very small area, thus obtaining very concentrated particles. For that purpose, collection is made directly on a nitrocellulose filter while a vacuum is established below the filter. By comparison with collection onto a microscope slide, the local concentration is increased about 1,000-fold. This efficient and simple system allows us to concentrate cell or cellular organelle populations for observation by microscopy of many objects in a single field, or to study the biological or biochemical properties of a small number of sorted particles.

Collodion↗

Localization of actin-related sequences by in situ hybridization to R-banded human chromosomes.

Using a cytoplasmic actin cDNA probe we have localized a number of actin sequences in the human genome using a novel in situ hybridization technique. Metaphase chromosomes treated to produce R-bands were directly annealed with 125I-labeled actin probe. Under these conditions many regions of the genome were apparently denatured enough to be capable of hybridizing with the probe. Most of the actin sites detected in prior experiments using chromosome preparations, which had been completely denatured, were recognized in this experiment. The major advantage of this method over standard in situ hybridization techniques is the marked increase in the resolution of subregional localization.

Actins↗

Mitochondrial myopathy with lactic acidosis and deficient activity of muscle succinate cytochrome-c-oxidoreductase.

A male infant had severe muscular hypotonia from birth. Recurrent vomiting with dehydration and severe metabolic acidosis complicated the course. Elevated lactate (up to 12.3 mmol/l; n less than 2), pyruvate (0.4 mmol/l; n less than 0.05) and alanine levels were found in serum with an abnormal lactate/pyruvate ratio (greater than 30; n less than 15). In urine the concentrations of lactate, pyruvate, alanine and of several intermediates of the citric acid cycle were increased. In muscle, numerous disseminated "ragged red fibres" were found by light microscopy; muscle fibres were found to contain subsarcolemmal aggregates of mitochondria, lipid droplets and glycogen by electromicroscopical methods. Moreover, mitochondria with a typical circular arrangement of cristae were noticed. In liver homogenates normal activities of pyruvate carboxylase and pyruvate dehydrogenase complex were found; in liver mitochondria also succinate-cytochrome-c-oxidoreductase activity was normal. However, in muscle no succinate-cytochrome-c-oxidoreductase activity was detectable. The patient became increasingly lethargic and died because of sepsis at 5 months of age.

Acidosis↗

Flow cytometry isolation and improved visualization of sorted mouse chromosomes. Purification of chromosomes X and ISO-1 from cell lines with Robertsonian translocations.

While analysis and sorting of human chromosomes by flow cytometry has been widely used, isolation of a pure mouse chromosome remains very difficult, since most murine chromosomes are quite similar in size. To overcome this problem, we have analysed mouse cell lines having either Robertsonian translocations or isochromosomes. The resulting metacentric chromosomes are very different in size and in morphology from normal mouse acrocentric chromosomes. These characteristics have been analysed by computer-monitored flow cytometry, facilitated by improvements in the chromosome extraction procedure. Signals characteristic of the iso-lq chromosome in cell line PCC4 azaR1, and of the normal X chromosome in the mouse strain 22CD have thus been obtained. These chromosomes have been sorted and can be easily recognized by fluorescence microscopy when collected onto serum-albumin-coated microscope slides. The technical modifications made, coupled with the existence of a great diversity of metacentric chromosomes resulting from Robertsonian translocations, should allow the purification of a number of different mouse chromosomes.

Animals↗

Antibody-induced conformational changes result in enhanced avidity of antibodies to different antigenic sites on the tick-borne encephalitis virus glycoprotein.

By the use of monoclonal antibodies we have recently defined eight distinct epitopes on the structural glycoprotein of tick-borne encephalitis (TBE) virus which differ with respect to location, function, or serological specificity (Heinz et al, Virology 126, 525-537, 1983). The present investigation reveals a complex network of interactions between antibodies directed against distinct nonoverlapping epitopes leading to enhanced binding of certain antibodies in the presence of bound second antibodies. The enhancement between antibody pairs can be either unidirectional or bidirectional. In addition, there are domains of predilection, which bind enhanceable antibodies (domain A) whereas others bind antibodies which preferentially induce enhancement (domain B). These domains represent structurally unrelated entities, domain A being sensitive to denaturation and fragmentation and domain B being resistant. Quantitative evaluation of binding data by Scatchard analysis revealed that the observed enhancement phenomenon is due to a two- to sixfold increase of antibody avidity. In the system described, enhancement of antibody binding is not dependent on antibody bivalency since it could also be demonstrated with purified Fab fragments acting either as enhanced or as enhancing antibody. It is therefore concluded that binding of antibodies to certain epitopes on the TBE virus glycoprotein induces conformational changes in distant parts of the molecule which can result in increased avidity of antibodies directed to conformationally changed epitopes. A possible explanation for the origin of this enhancement phenomenon is presented.

Antibodies, Monoclonal↗

Cytogenetic studies on acute nonlymphocytic leukemias following polycythemia vera.

Chromosome studies were performed on 15 patients suffering from acute nonlymphocytic leukemia (ANLL) and in one patient in a preleukemic state following polycythemia vera (PV). Clonal chromosome abnormalities that were present in all cases were clearly nonrandom and involved chromosomes #1, #5, #7, #8, #9, #11, and #21. A subdivision of ANLL into two categories occurring in the course of PV is proposed from the clinical, hematologic, and cytogenetic data: one resembling de novo ANLL with rapid initial evolution, easy classification into one group of the FAB nomenclature, and simple chromosome abnormalities; the other resembling induced leukemia, often with more progressive initial evolution, difficulty or impossibility of classification into one group of the FAB nomenclature, and complex chromosome abnormalities. The consequences for the commitment level of progenitor cell from which the leukemic clones originate are discussed.

Acute Disease↗

Chromosome studies in polycythemia vera patients.

One hundred thirty-five polycythemia vera (PV) patients (30 untreated by chemotherapy and 105 treated) were studied cytogenetically . The incidence of clonal chromosomal abnormalities was 20.7% (28 patients in nonleukemic phase). The incidence of 20q - was 3.7% (5 patients). The presence of cytogenetically abnormal clones did not allow prediction of the evolution of the disease. In a few cases, abnormal clones disappeared at the time of later studies. Although nonrandom, the majority of clonal chromosomal abnormalities are believed to be secondary events in PV patients.

Aged↗

A third example of haemolytic auto-anti-Vel.

A non-transfused, 43 year old Caucasian female presented with acute haemolytic anaemia and splenomegaly. Sections of bone marrow showed erythroid hyperplasia. The patient's red blood cells gave a negative reaction with polyspecific antiglobulin serum, but a positive reaction with specific anti-IgM. A heat eluate prepared from her red cells showed anti-Vel specificity. Her serum agglutinated only Vel-positive cells including her own. All papain pre-treated red cells including her own and Vel-negative cells were completely haemolysed at 37 degrees C. The percentage of haemolysis of Vel-positive cells was greater than that of Vel-negative cells.

Adult↗

Induction of interferon gamma in mouse spleen cells by culture supernatants of mycoplasma arthritidis.

Factor(s) in the supernatant of cultured mycoplasma arthritidis (MAS) represented potent inducers of interferon (IFN) in cultures of mouse spleen cells. Responding mouse strains included A/J, BALB/c, CBA, C3H, and DBA/2 whereas spleen cells of C57BL/6 and C57BL/10 mice were nonresponders. Interferon production was controlled by the I region of the H-2 locus. Treatment of CBA spleen cells by anti-thy-1 antibody plus C abolished IFN production. A similar effect was seen when CBA spleen cells were freed of macrophages by passage through Sephadex G-10 columns. Pure macrophages themselves, however, were not producing IFN when treated by MAS. Macrophage-depleted CBA spleen cells could be reconstituted to produce IFN by the addition of 2-ME. Interferon induction by CBA spleen cells was independent of lymphoproliferation as evidenced by experiments utilizing mitomycin C. The IFN induced by MAS represented IFN gamma (gamma), since it was acid-labile and neutralized by a specific antiserum.

Animals↗

Immunogenicity of tick-borne encephalitis virus glycoprotein fragments: epitope-specific analysis of the antibody response.

After digestion with trypsin, alpha-chymotrypsin, or chemical cleavage using CNBr, fragments of the tick-borne encephalitis (TBE) virus glycoprotein were isolated which retained their reactivity with neutralizing monoclonal antibodies defining a denaturation-resistant antigenic domain. Upon immunization of mice, these fragments induced antibodies reactive with the immunizing peptide, the denatured glycoprotein and the native glycoprotein as a constituent of the whole virus. The immune sera revealed the same properties as the monoclonal antibodies that were used to select the fragments for immunization: neutralizing activity; haemagglutination-inhibiting activity; blocking of the binding of antibodies used for selection; enhancement of the binding of other monoclonal antibodies defining a denaturation-sensitive antigenic domain. It was shown that the natural immune response against certain functionally important, denaturation-resistant immunogenic domains on the native protein can be closely mimicked by immunization with defined protein fragments. Antigenic sites present on these fragments may therefore represent essential constituents of a synthetic vaccine. The fine specificities of antibody populations in anti-peptide or anti-protein immune sera were analysed on the basis of single antigenic determinants by blocking assays using radiolabelled monoclonal antibodies that define eight distinct epitopes on the TBE virus glycoprotein. Quantitative differences in the blocking of certain monoclonal antibodies were also observed between human convalescent sera. The establishment of such blocking profiles using a panel of well-characterized monoclonal antibodies may represent a general method for dissecting the specificities of antibody populations present in polyclonal immune sera and could allow investigations on determinant-restricted differences of immune responses and its possible implications for the course of the disease.

Animals↗

Malignant lymphomas with band 8q24 chromosome abnormality: a morphologic continuum extending from Burkitt's to immunoblastic lymphoma.

Chromosome abnormality involving band 8q24 is present in the malignant cells in virtually all 'Burkitt's' type malignancies. t(8;14)(q24;q32) translocations have nevertheless been found in certain cases of malignant lymphomas (ML) described as 'small non-cleaved non-Burkitt', 'immunoblastic' or 'histiocytic'. With a view to comparing objectively the histological picture of such lymphomas, we undertook morphometric analysis of seven cases of diffuse ML classed as 'Burkitt's' (three cases), 'immunoblastic' (two cases), 'small non-cleaved non-Burkitt' (one case) and 'large-cell lymphoma' (one case), all exhibiting band 8q24 rearrangement arising from various translocations. Our study substantiates the view that the histologic picture of MLs with 8q24 anomaly fits into a morphological continuum containing 'Burkitt's', 'small non-cleaved non-Burkitt' and 'immunoblastic' lymphomas.

Adult↗