Comparative analysis of high-resolution chromosome techniques for leukemic bone marrows

High-resolution direct and synchronization culture techniques for chromosome analysis of leukemic bone marrow cells can now be utilized. In this article, three different techniques are quantitatively compared for their consistency in successful cytogenetic analysis, reliability in the detection of clones with chromosomal abnormalities, and usefulness for the precise delineation of break points involved in structural chromosomal rearrangements. Bone marrow samples from 15 consecutive patients with acute nonlymphocytic leukemia (ANLL) were studied using an improved direct technique, amethopterin cell synchronization with thymidine release, and amethopterin cell synchronization with bromodeoxyuridine (BrdU) release. The results obtained with the amethopterin cell synchronization technique and thymidine release suggest that it should be the method of choice in the detection of chromosome defects in bone marrow of patients with ANLL.     

S-100 protein immunostaining in the differential diagnosis of chondroblastoma

In an effort to investigate the utility of immunostaining for S-100 protein in the differential diagnosis of chondroblastoma, the expression of S-100 protein in nine chondroblastomas was compared with that in six giant cell tumors, six aneurysmal bone cysts, four giant cell reparative granulomas, six cases of fibrous dysplasia, two cases of osteitis fibrosa cystica, two nonossifying fibromas, and one clear cell chondrosarcoma. Five enchondromas, three typical chondrosarcomas, and one mesenchymal chondrosarcoma were also included as control tumors. The proliferating stromal cells in seven of the nine chondroblastomas stained for S-100 protein, as did the lacunar chondrocytes in all of the enchondromas and chondrosarcomas and rare stromal cells in the clear cell chondrosarcoma. In contrast, none of the other tumefactive bone lesions included in this study demonstrated S-100 protein immunoreactivity. These results suggest that immunohistochemical assessment of S-100 protein may be a method for diagnostically separating chondroblastoma from pathologic entities that could be histologically confused with it in the presence of limited biopsy material. However, clear cell chondrosarcoma would appear to represent an exception to this general statement.     

Discontinuation of antipsychotic drug therapy

When an antipsychotic medication is discontinued, the possible withdrawal symptoms include psychotic relapse, medical complaints, and dyskinetic movements. Psychotic relapse, if it occurs, may be delayed for months or years and can be detected by close patient follow-up. Medical symptoms, such as nausea, Vomiting, and dizziness, are time-limited. Tardive dyskinesia may appear during or after drug withdrawal, but this problem is not necessarily permanent. To minimize symptoms, antipsychotic drugs should be tapered gradually, and patients on an antiparkinsonism medication should continue it for at least a week after the antipsychotic drug is stopped.     

High resolution chromosomes of the t(9;22) positive leukemias

A combined bromodeoxyuridine (BrdU) and actinomycin D (AMD) treatment of methotrexate (MTX) synchronized bone marrow cells from four patients with chronic myelogenous leukemia (CML), and two each with acute lymphocytic (ALL) and acute nonlymphocytic (ANLL) leukemia were used to define the breakpoints involved in the t(9;22) found in subgroups of these three disorders. An additional 20 patients with CML were studied with the MTX technique alone. All cases showed the same breakpoints at subbands 9q34.1 and 22q11.2. In CML, it was possible to further define the breakpoint of chromosome 22 to subband q11.21.     

Nomenclature for high resolution human chromosomes

The recent development of high resolution chromosome banding techniques is having a major impact in the study of cancer, mental retardation, birth defects, chromosome structure, gene mapping, and evolution. To establish a standard of communication, a nomenclature for long, finely banded chromosomes is proposed. The system strictly adheres to the original guidelines of the Paris Conference, incorporates information derived from new high resolution G-, Q-, and R-banding techniques, accurately portrays the process of band fusion during chromosome condensation, and assigns a unique number to every new chromosome band (subband) observed, regardless of stage of chromosome condensation and total number of bands visualized.     

Early detection and specificity analysis of human cytolytic T lymphocyte (CTL) colonies generated in soft agarose culture: a potential assay for definition of CTL defined (CD) determinants

In this communication we describe and early, large-scale screening assay for the detection of colonies with varied cytolytic specificity. The colonies are generated in soft agarose culture from day 3 mixed lymphocyte culture (MLC) alloactivated cells. A cell mediated lympholysis (CML) assay utilizing as few as 500 target cells has made it possible to prescreen for cytolytic T lymphocyte (CTL) colonies and to test for antigen specificities as early as 11 and 14 days, respectively, after MLC priming. Large numbers of colonies, from 80 to over 150, have been prescreened against a specific sensitizing target cell, and as many as 30 CTL colonies have been simultaneously tested against a panel of multiple targets carrying defined HLA-A, -B, -C, -DR antigens to evaluate antigen specificity. All CTL colonies are lytic against the specific sensitizing target cell and do not lyse the target autologous to the responder. Some are found to be operationally specific in that they lyse only those target cells which share HLA serologically defined (SD) antigens with the sensitizing cells, and other show cytolytic patterns which are not correlated with known HLA-SD antigens. These observations support, at a much finer level of analysis, the possible distinction between SD and CTL defined (CD) determinants.     

Improving the effectiveness of psychiatric consultation

Drawing on their studies of consultation outcomes, the authors propose ways to improve consultees’ concordance with psychiatric consultants’ recommendations and diagnoses. Specific steps to improve outcome are presented. These include identification of cases at high risk for nonconcordance, attention to the form and substance of the consultation note, and monitoring the consultees’ responses to consultants’ recommendations.     

Banded chromosome analysis in patients with treatment-associated acute nonlymphocytic leukemia

We have analyzed G-banded metaphase chromosomes from 20 patients with treatment-associated acute nonlymphocytic leukemia (t-ANLL). Nine patients were previously treated for hematologic malignancies and 11 for solid tumors. The interval from initial therapy to t-ANLL ranged from 35 to 182 mo (median 75.5 mo). Medial age at diagnosis of t-ANLL was 58.5 years. Clonal chromosome abnormalities were found in 19 patients (95%). Loss or partial deletion of the long arm of chromosomes #5 and/or #7 were most common, occurring in nine patients. These abnormalities were associated with hypodiploid complex karyotypes. Other nonrandom abnormalities recurring among karyotypes with abnormalities of chromosome #5 included loss of one #18, partial deletion of the long arm of chromosome #2, ring chromosomes, and a Philadelphia (Ph1) chromosome. We also identified a group of five patients whose only karyotypic abnormality was addition of whole chromosomes. The remaining five patients had other karyotypic abnormalities, the most common of which were structural rearrangements in a pseudodiploid clone. Combined data from our study and the three previously published large series of patients with t-ANLL studied with banding suggest a relationship between karyotype and intensity of prior therapy, with abnormalities of chromosomes #5 and #7 occurring more often in the intensively treated patients.     

Retinoblastoma and its association with a deletion in chromosome #13: A Survey using high-resolution chromosome techniques

To our knowledge, 20 cases of retinoblastoma associated with a chromosome #13 aberration have been reported. The present study utilized high-resolution prophase banding analysis of 12 additional retinoblastoma patients to determine the occurrence of chromosome aberrations and identify consistently associated clinical abnormalities. Six male and six female patients were studied representing seven cases of bilateral and five cases of unilateral retinoblastoma. One case of unilateral and two cases of bilateral retinoblastoma had detectable cytogenetic abnormalities, all involving an interstitial deletion of 13q14 on the long arm of one chromosome #13. In all five unilateral cases the tumor manifested in the left orbit, and in all seven bilateral cases the left eye was at a more malignantly advanced stage than the right eye. All three cases with a chromosome abnormality had varying degrees of developmental and/or mental retardation, along with at least one other congenital abnormality. In addition to the 12 cases of retinoblastoma, a patient with severe ophthalmologic abnormalities and mild congenital anomalies was studied by the prophase banding technique and found to be partially trisomic for the q13–14 banding regions of chromosome #13. These results further confirm the association of the 13q14 region with gene loci for optic development and indicate that cytogenetic abnormalities may occur even more frequently in retinoblastoma than indicated by the small number of cases reported in the literature.     

Leukocyte chemotaxis under agarose: manipulations of serum and plasma before incorporation into agarose can influence cell movement

In the migration under agarose assay for measurement of polymorphonuclear leukocyte chemotaxis and random migration, the agarose gel ordinarily contains 10% human serum or plasma. Influences of serum and plasma on cell movement using this assay were defined. Utilizing plasma as protein source allowed more random migration and chemotaxis than when serum was used. Heat inactivation of both serum and plasma decreased random migration and chemotaxis. If heating at 56 degrees C was prolonged beyond 15--30 min, leukocyte movement was augmented. Excessive concentrations of platelets or platelet products, or heparin in agarose, also augmented migration, but storage of plasma or sera at various temperatures for 24 h did not appreciably affect migration. Large amounts of hemoglobin in agarose decreased chemotaxis, but the effect of trace amounts of hemolysis was negligible. Handling plasma or sera in standard ways prior to use in the under agarose assay may make this assay more reproducible and sensitive.     

Recurrent chromosomal defects are found in most patients with acute nonlymphocytic leukemia

Using a methotrexate cell synchronization technique, we have successfully studied chromosome preparations of bone marrow biopsies from 49 of 51 patients with acute nonlymphocytic leukemia (ANLL). Clonal chromosomal abnormalities were detected in 46 patients, and one of eight types of recurrent chromosomal aberrations were found in 31 patients. One of these types represents a new chromosome defect [inv(16)(p13.1q22)], found in four patients with ANLL-M2, -M4. Chromosome preparations from cultured lymphocytes of two of the four patients were analyzed for the presence of chromosome fragile sites. Both cases showed a frequent break at band 16q22, an intriguing finding that suggests a possible correlation between a fragile site and predisposition to chromosomal rearrangement in human neoplasia.     

Prognostic significance of the Philadelphia chromosome in acute lymphocytic leukemia

Prognostic factors for acute lymphocytic leukemia (ALL), including initial white blood count (WBC), age at diagnosis, sex, cytology, lymphocyte surface markers, and glucocorticoid receptors, have been evaluated in 12 patients presenting with ALL and the Philadelphia chromosome (Ph¹+ ALL). Ph¹+ ALL patients had a median initial WBC of 89 × 10⁹ liter, a median age of 23 years and a male: female ratio of 1:1. The cytology of the blasts was L1 in 50% and L2 in 50%. The blasts in all cases studied were characteristic of non-T, non-B, common ALL, had high terminal deoxynucleotidyl transference levels, and low numbers of glucocorticoid receptors. The prognostic factors in these Ph¹+ ALL patients were compared with those of a concurrent group of adults with Ph¹? ALL. Patients with Ph¹+ ALL more commonly had unfavorable prognostic features (high WBC, older age, male sex, L2 cytology, and low glucocorticoid receptors). They also had fewer complete remissions (58 vs. 100%, p=0.07), shorter remission durations (median 10 vs. 44+ months, p=0.001) and shorter survival times (median 11 vs. 48.5+ months, p=0.00008). However, even when patients with similar prognostic features were compared, those with the Ph¹ had fewer and shorter remissions and shorter survival times than ALL patients without the Ph¹. These data suggest that the Ph¹ is an independent unfavorable prognostic factor in adults with ALL.     

Recurrent chromosomal defects are found in most patients with non-Hodgkin's-lymphoma

Using methotrexate cell synchronization, we successfully analyzed chromosomal preparations of 40 lymph node biopsies and one bone marrow sample from 44 patients with non-Hodgkin's, non-Burkitt's lymphoma. All of the 41 patients successfully analyzed showed clonal chromosomal abnormalities. In 25 of the 41 (61%), the defects were found to be consistent with (A) a deletion 6q in five of seven patients with diffuse large cell lymphoma, (B) a t(11;14), a del 11q, or a +12 in seven of nine patients with small cell lymphocytic lymphoma, and (C) a t(14;18) in 12 of 15 patients with follicular lymphoma (small cleaved and mixed small and large cleaved) and in a single case of diffuse large cell lymphoma. In three patients with small cell lymphocytic lymphoma whose biopsies exhibited a t(11;14), lymphocytes were cultured and chromosomes examined for the presence of fragile sites. In two, frequent breaks at band 11q13.3 were observed. Such findings suggest a possible relationship between a fragile site and a predisposition to a specific chromosomal rearrangement in human neoplasia.     

Hepatic veno-occlusive disease after high-dose mitomycin C and autologous bone marrow transplantation therapy

Three cases of veno-occlusive disease of the liver were diagnosed in four autopsied patients who had received high-dose mitomycin C therapy followed by autologous bone marrow transplantation, and the pathologic finding are reported. Review of 27 liver, examined post mortem, of patients receiving other high-dose chemotherapeutic regimens, 15 of them with subsequent autologous bone marrow transplantation, revealed no evidence of veno-occlusive disease. Veno-occlusive disease may now become a dose-limiting factor in the use of the combined high-dose mitomycin C-bone marrow transplantation therapy. Attention is also drawn to the increasing number of veno-occlusive disease cases being reported in associated with alkylating agents.     

Preserved human remains from the southern region of the North American continent: report of autopsy findings

Autopsies were performed in six naturally mummified bodies from the southern area of North America. The cause of death was established with reasonable certainty in two (abdominal stab wound and sand pneumoconiosis); was somewhat speculative in one (atelectasis secondary to bronchial obstruction by an aspirated tooth); and was undetermined in three. The value of such studies is discussed, and the synergistic effect of interdisciplinary contributions is emphasized.