Biomedical subjects

J M Trujillo

Publications and source records attributed to J M Trujillo.

At least 109 records · Page 6Linked to original sources

PubMed1985

Suppression of clonal evolution in two chronic myelogenous leukaemia patients treated with leucocyte interferon.

Two patients with Philadelphia chromosome-positive (Ph1+) chronic myelogenous leukaemia (CML) were treated with human leucocyte interferon (HuIFN-alpha). Karyotypic changes in addition to the Ph1 chromosome developed in these patients before the start of HuIFN-alpha treatment. In one patient the administration of HuIFN-alpha resulted in clinical haematological remission and stable suppression of the secondary Ph1 clone. The second patient was in myeloid blastic crisis when given HuIFN-alpha. While she was receiving HuIFN-alpha, suppression of the blast cell population in the bone marrow occurred. The subsequent cytogenetic changes included a near-complete suppression of a secondary Ph 1 clone of cells carrying a deletion in the short arm of chromosome 7 and partial population of the bone marrow with primary Ph1 clone. These observations suggest a potential role for interferons in altering the progressive course of CML.

Adult↗

PubMed1985

E rosette-positive agar colonies containing the Philadelphia chromosome in chronic myeloid leukaemia.

In an attempt to document possible T-cell involvement in chronic myeloid leukaemia, E rosette-positive colonies (ERPC) were grown in agar culture using a T-cell-conditioned medium. Colonies were grown from whole mononuclear cells (WMN), nonadherent E rosette-positive (NAT+) and nonadherent E rosette-negative (NAT-) cells. Cells collected from the colonies after 10 d in culture were 99-100% E rosette-positive. 8 metaphases were obtained from both NAT+ and NAT- ERPCs. In NAT- ERPCs, 5 out of 8 metaphases were positive for the Philadelphia (Ph1) chromosome as compared to 1 Ph1 positive out of 8 metaphases in the NAT+ ERPCs. These results suggest that, at least in this particular patient we studied, a subpopulation of E rosette-positive cells derived from the NAT- cell fraction express the Ph1 chromosome.

Adult↗

PubMed1984

ARH-77, an established human IgG-producing myeloma cell line. I. Morphology, B-cell phenotypic marker profile, and expression of Epstein-Barr virus.

Monolayer cultures of ARH-77 cells, a human myeloma cell line propagated in vitro, display a variety of morphologic entities ranging from small lymphocytes to classic plasma cells. The cells show intense pyronin and periodic acid-Schiff affinity but are negative for colloidal iron, sudan black, and naphtol AS-D chloroacetate esterase. The cells exhibit phenotypic markers pertaining to each stage of the B-cell lineage. They fail to display sheep erythrocyte and bovine erythrocyte-IgG antibody complex rosettes, common acute lymphocytic leukemia (ALL) antigens and T-cell antigens, but most cells display surface complement receptors, Ia-like antigens, and surface and intracytoplasmic Ig. Monoclonal antibodies were negative for T-antigens, myelomonocytic cell antigens, leukemia-associated antigens, and BA-1 and OKT-10 antigens. However, 100% of the cells were positive with OKT-9 and B3/25 antibodies that are specific for transferrin receptors. About 50% to 80% of the cells were positive for surface membrane immunoglobulin (kappa IgG) and about 10% to 50% for cytoplasmic immunoglobulin (kappa IgG). Virtually all cells were positive when tested for nuclear Epstein-Barr virus antigens.

Adult↗

PubMed1984

ARH-77, an established human IgG-producing myeloma cell line. II. Growth kinetics, clonogenic capacity, chalone production, xenogeneic transplantations, and response to melphalan.

The growth curve of monolayer cultures of ARH-77 cells, a human myeloma cell line propagated in vitro, is represented by an everbending curve on a semilogarithmic plot; however, the curve can be fitted by a straight line on a linear-linear plot. This unusual growth pattern suggests that, instead of a fixed proportion of the population, a fixed number of ARH-77 cells divide per unit time. The following are cell cycle transit time parameters calculated from percent labeled mitosis experiments: TG1, 10.0 +/- 3.5 hours; Ts, 14.3 +/- 2.3 hours; TG2, 4.3 hours; TM, 1.4 +/- 1.3 hours; and Tc, 30.0 +/- 6.1 hours. For cells exposed continuously to 3H-thymidine the values are: growth fraction, 67%; TG1, 6.5 hours; Ts, 13.0 hours; and TG2 + M, 3.0 hours. The average doubling time is 4.6 days (range, 3.8-4.7 days); after about 10 to 15 days in culture, the growth rate of freshly passaged cells declines markedly, as reflected by a growth curve with a much shallower slope. The changes are accompanied by a marked decline in the labeling index from 41.3% (range, 28.9%-53.7%) during the first 3 days of culture to less than 5% measured on day 21. Flow cytometry for DNA content-dependent cell cycle compartment distribution demonstrates an obvious decline in the proportion of S-phase cells and a marked accumulation of G2 phase cells as the cultures age. When the supernatant medium of ARH-77 cells grown for 10 days is replaced by fresh medium, a new burst of vigorous cellular growth is observed with a curve slope similar to that observed during the first 5 days of culture. If the 10-day-old supernatant medium is used to set up cultures with freshly harvested ARH-77 cells, their growth curve resembles that of 10-day-old cultures. However, this supernatant medium induces no decrease in the growth rate of other human tumor cells, suggesting that inhibition of cellular growth does not result from exhaustion of nutrients, but that ARH-77 cells produce a molecular mediator that specifically inhibits the growth of these cells. ARH-77 cells could be synchronized with a single treatment of 3 or 5 mM thymidine; (dThd) and cloning efficiency was 2% to 4% in a double-layer soft agar assay. Treatment for 1 hour with increasing concentrations of melphalan produced a threshold exponential survival curve (Dq = 0.45 microgram/ml and D0 = 0.35 microgram/ml, 1 hour).(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

PubMed1984

Evidence for extensive intermolecular disulfide bonds of the proteins of non-ionic detergent high-salt-resistant skeletons of normal lymphocytes, and the altered structure in leukemia.

Normal and leukemic lymphocytes were subjected to lysis with a non-ionic detergent and 2.0 M NaCl. The intact nuclear DNA and the associated molecules resulting from these lyses (the nucleoids) were then separated from DNA-dissociated molecules and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Most of the proteins undissociable by 2.0 M NaCl from normal lymphocyte nuclear DNA or from DNA-associated molecules appear to be intermolecularly S-S linked. A distinct set of these proteins was either absent or only present in greatly decreased amounts in association with the leukemic lymphocyte nuclear DNA exposed to 2.0 M NaCl. On the other hand, the opposite relationship was found for a an MW 71,000 protein with leukemic lymphocytes of every patient studied regardless of the subtype of leukemia. Conversion of normal quiescent lymphocytes to cycling cells by lectin stimulation rendered them similar to leukemic cells in some respects but not in all. Our results show an altered nuclo(cyto)skeletal structure in leukemic cells; in general the proteins involved appear to be sulfhydryl proteins whose redox states and/or DNA association properties are modified by the neoplastic transformation of lymphocytes.

Blood Proteins↗

PubMed1984

Cultured human tumour cells may be arrested in all stages of the cycle during stationary phase: demonstration of quiescent cells in G1, S and G2 phase.

Six human colon carcinoma cell lines were induced to enter stationary phase of growth by nutrient deprivation and cell crowding. Growth kinetics parameters (cell number, flow cytometric analysis of DNA distribution, and labelling and mitotic indices) were measured sequentially for all lines during the various stages of in vitro growth. Our results demonstrated that a substantial fraction of cells (9-18%) were located in G2 phase when they changed from an exponential to a stationary mode of growth. Moreover, a large number of cells in stationary phase of growth had an S-phase DNA content, as determined by flow cytometry, but failed to incorporate radioactive DNA precursors (up to 15-fold difference). To substantiate these findings, cells in stationary phase of growth were induced to enter exponential growth by re-seeding in fresh medium at a lower density. Subsequently observed changes in DNA-compartment distribution, and in labelling and mitotic indices were those expected from cells that had been arrested at different stages of the cycle during their previous stationary phase. Thus, the non-proliferating quiescent state (Q), traditionally located 'somewhere' in G1 phase, appears to be composed also of cells that can be arrested at other stages of the cycle (Qs and QG2). Although the proportion of such cells is rather small, their contribution to the growth kinetics behaviour of human in vivo tumours will become apparent following 'recruiting' or 'synchronizing' clinical manoeuvres and will prevent the formation of a clear-cut wave of synchronized cells.

Cell Cycle↗

PubMed1984

Use of Imugard IG500 filters for preparation of leukocyte-poor blood for cancer patients.

In order to determine the efficiency of Imugard IG500 filters in producing leukocyte-poor blood (LPB), 109 units of red cell concentrates were processed using cold (4 degrees C) or room temperature saline solution without the application of pressure to the blood bag. The results obtained in terms of red blood cell recovery, white blood cell removal, residual white blood cells, and free hemoglobin in the supernatant of the filtered units are comparable to those processed with the IBM 2991. LPB processed without pressure application to the blood bag and room temperature saline solution can be rapidly, efficiently and safely prepared in any size blood bank/transfusion service. This technique is definitely cost-saving in terms of equipment, software, space and personnel training.

Blood Group Incompatibility↗

PubMed1984

Cellular discriminants for a biological classification of human colon carcinoma.

We categorized established human colon carcinoma cell lines into three biological groups. Our studies were performed on six colorectal cancer lines representing the proposed three groups. Group I consisted of two lines designated LoVo and SW 48; Group II comprised two lines called SW 480 and SW 620; and Group III was represented by lines SW 403 and SW 1116. Group I consisted of the most differentiated cells. This differentiation encompassed morphological markers, gland and signet ring formation, and ciliary development. The outstanding morphological characteristic of Group III was the development of numerous multinucleated giant cells. The range and modal chromosome number increased from Group I to Group III, a change reflected by the higher DNA content of these cells as measured by flow cytometry. Carcinoembryonic antigen synthesis was maximal for Group III and virtually absent for Group II. The number of clonogenic cells decreased from Group I to Group III, while the proportion of nonproliferating cells calculated both by experiments using continuous labeling with tritiated thymidine, and by the primer-available alpha-DNA polymerase index, increased from Group I through Group III. Another important cytokinetic difference was that Group I had an exponential cell cycle stage distribution not seen for the other groups. Cells in Group I were easily propagated in athymic (nude) rats by s.c. injection; cells in Group II injected s.c. grew for about 30 days and then regressed spontaneously. Cells in Group III could only be grown when inoculated intracerebrally. Thus, our studies have now confirmed and extended the hypothesis that cultured human colorectal carcinomas can be separated into at least three groups on the basis of morphological differentiation, chromatin distribution, carcinoembryonic antigen production, cytokinetic properties, and xenograft propagation. Perhaps this classification is just the tip of the iceberg, and future studies will determine the existence of additional groups or subgroups on the basis of other markers. However, at present it appears established that malignant cells with a common histological origin in the gut express their phenotypic potential in a sufficiently discrete manner as to permit their classification into distinct biological groups. Thus, the stage is set for extrapolating this in vitro classification for an in vivo segregation of human colorectal tumors into categories with specific properties and diverse prognosis.

Animals↗

PubMed1984

Association of circulating immune complexes with cytogenetic abnormalities but not with prognosis in acute nonlymphocytic leukemia.

Sera from 91 adult acute nonlymphocytic leukemia patients were tested for circulating immune complexes by the C1q binding test. In 42 patients tested prior to the initiation of therapy, C1q binding activity was inversely correlated with leukemic infiltrate of the bone marrow (rs = -0.34, p less than 0.02), but it was not related to peripheral white blood cell count or presence of infection. Patients with abnormal C1q binding activities (greater than 6%) were more likely to have cytogenetic abnormalities in their bone marrow (p = 0.02); the most frequent abnormality was an extra No. 8 chromosome. The median survival of 8 patients with abnormal values was 1.5 months as compared to 7.8 months in 34 patients with normal values. However, the C1q binding test did not identify patients likely to achieve remission; and after remission was achieved, it did not identify those likely to relapse. Differences between these findings and those recently published by other investigators are discussed.

Acute Disease↗

PubMed1983

Comparative cytotoxicity of bisantrene, mitoxantrone, ametantrone, dihydroxyanthracenedione, dihydroxyanthracenedione diacetate, and doxorubicin on human cells in vitro.

The cytotoxic efficacies of several substituted anthraquinones, ametantrone, dihydroxyanthracenedione, dihydroxyanthracenedione diacetate, mitoxantrone, bisantrene, and doxorubicin, were evaluated on an established human colon adenocarcinoma cell line by the method of inhibition of colony formation. The concentration-dependent survival curve following treatment for 1 hr was biphasic exponential for all agents. At concentrations below 1 microgram/ml, mitoxantrone was about twice as active as both hydroxyl-substituted anthracenediones and doxorubicin, about 14 times more efficacious than ametantrone, and about 22 times more powerful than bisantrene. At higher concentrations, these differences in efficacy became even more pronounced. Treatment in stationary phase decreased the lethal efficacy of doxorubicin but not that of the other agents. No recovery of potentially lethal or sublethal damage was noted for any agent, but for anthracenedione derivatives, there was a small but statistically significant increase in cell kill during fractionated exposure. Continuous treatment with mitoxantrone or bisantrene resulted in marked degrees of cell killing, reaching 99.95 and 99.5%, respectively, after 24 hr. For doxorubicin, cell kill efficacy declined after 4 hr. Mitoxantrone was 10-fold more active on cells in G2 phase than on those in mid- to late-S phase. Sensitivity in G1 phase was intermediate. Thus, mitoxantrone appears as the most active compound while bisantrene and ametantrone are the least active agents. The cytotoxic efficacy of bisantrene increases during prolonged continuous exposure, while that of mitoxantrone increases in fractionated administration. These characteristics could be exploited in clinical strategies designed to improve the performance of these agents.

Anthracenes↗

PubMed1982

A comparative study of the effectiveness of different mitogens for karyotypic analysis in chronic B cell leukemia.

In order best to study the karyotypic abnormalities of nine patients with chronic lymphocytic leukemia of B cell origin, we used a combination of T and/or B cell mitogens so as to achieve an optimal proliferative response. As assessed by DNA synthesis, and subsequent cytogenetic study, pokeweed mitogen (PWM) and protein A Sepharose (PROT A) appeared to hold promise as potent stimulators of neoplastic B lymphocytes. Lipopolysaccharide from E. coli (LPS) proved to be an ineffective mitogen. Phytohemagglutinin (PHA), while producing the highest stimulation indices by DNA assay, appeared to induce predominantly diploid metaphases. Pseudodiploid clonal abnormalities were noted in three out of nine patients, while another four patients showed random structural abnormalities, including translocations, which were indicative of damage. Presence of the A33 antigen, as determined by HLA typing, was noted in five out of nine patients. The increased frequency of this antigen in patients with chronic B cell leukemia is discussed.

Adult↗

PubMed1982

Serum lactate dehydrogenase isoenzyme 1 in patients with advanced testicular cancer.

Abnormal levels of serum lactic dehydrogenase-1 (LD-1) activity have been observed in 81% (34/42) of patients with stage III germ cell malignancy of the testis. The criteria for evaluation the electrophoretic isoenzyme patterns of these patients were, as follows: For criterion 1 elevations the LD-1 value in absolute units was greater than 52.0 U/I with the LD-1/Total LD ratio greater than 37.2%. Criterion 2 elevations had absolute values of LD-1 less than 52.0 U/I, but the LD-1/Total LD ratio was greater than 37.2%. For criterion 3 elevations of LD-1, the absolute value was greater than 52 U/I and the LD-1/Total LD ratio was less than 37.2% activity with the LD-5/LD-1 ratio less than 0.5; or when the LD-5/LD-1 ratio was greater than 0.5 but the LD-1 is equal to or greater than the LD-2. The frequency of LD-1 elevation correlated well with the extent of the disease (stage II-B-1 and 2, 50%; stage III-B-3, 86%; stage III-B-4, 91%; stage III-B-5, 93%). LD-1 elevation occurred in groups I, II, IV and V histopathologic cell types (Dixon and Moore Classification) and there did not appear to be any correlation between the histologic cell type and the frequency of elevation of LD-1. Interpretation of LD-1 activity only on the basis of its relative ratio to the total LD value (criterion 1 and 2) identified a total of 28 patients (67%). A criterion 3 elevation was demonstrated in 6 (14%) additional patients. All patients with persistent elevations or recurrent elevations of LD-1 have shown progressive or recurrent disease and patients with no clinical evidence of disease have demonstrated normal LD-1 values. In those patients with elevated LD-1 activity, serial measurements of serum. LD-1 isoenzyme reflect the response of the patient to therapy.

Choriocarcinoma↗

PubMed1982

Eight-parameter automated hematology analyzers: comparison of two flow cytometric systems.

The performance of two high-speed 8-parameter automated hematology analyzers (Coulter Counter S-plus and ELT-8) were compared with that of reference instruments (Coulter Counter S Sr and Technicon Autocounter). The precision, linearity, and lack of carry-over of both instruments were superior over that of existing equipment. The especially noteworthy feature of the instruments was their excellent performance in the range of extreme values of both white blood cells and platelets. This enhanced performance, and the fact that all presently known relevant hematologic parameters can be measured by a single instrument on a single sample, make the contribution of these two flow cytometric-based instruments a significant advance in the field of automated laboratory medicine.

Autoanalysis↗

PubMed1982

A micro-assay for estrogen receptor in breast tumor with use of 125I-labeled estradiol.

The concentration of estrogen receptor protein in breast-tumor tissue is generally expressed in units of femtomoles of estradiol bound by the receptor per milligram of cytosol protein. The sensitivity of the estrogen receptor radioligand assay is therefore related to the specific activity of the steroid label used for the binding assay, the amount of the receptor protein in the volume of cytosol used, and the protein concentration in the cytosol. In this paper, we discuss factors affecting the sensitivity of the estrogen receptor assay and present various approaches for optimizing the assay. We also describe a procedure that involves a radioiodinated estradiol of high specific activity, a micro-technique for preparing tumor cytosol, and a micro-assay procedure with which the estrogen receptor protein can be measured in as little as 50 mg (wet weight) of tissue.

Breast Neoplasms↗

PubMed1982

Growth characteristics of human colonic adenocarcinomas propagated in the Rowett athymic rat.

Two human colonic adenocarcinoma cell lines were propagated in the Rowett athymic rat. Line LoVo displayed a well-differentiated morphology and exhibited progressive growth over a 70-day observation period with a doubling time of 8.5 days throughout. No metastatic involvement was noted. Xenografts of SW 620 cells were undifferentiated and highly necrotic. These tumors grew progressively for approximately 30 days with a doubling time of 5.5 days, but over 90% of the animals exhibited a spontaneous regression with a mean time to total regression of 51 +/- 3 (S.E.) days. Animals which had rejected the SW 620 xenografts would not support the growth of either the SW 620 or LoVo xenografts when challenged a second time with inocula producing a 100% tumor incidence in control rats. No metastatic involvement was noted for the SW 620 xenografts, but the tumors frequently invaded the underlying musculature and protruded into the peritoneal cavity without producing ascites or tumor nodules at sites distant from the primary.

Adenocarcinoma↗

PubMed1981

Clinical implications of cytogenetic abnormalities in acute nonlymphocytic leukemia.

Banding techniques have been applied to the study of the karyotypic alterations associated with acute nonlymphocytic leukemia (ANLL). These studies have identified nonrandom changes which preferentially involve certain chromosomal elements. Correlation of these chromosomal aberrations with the clinical ahd hematological findings indicates the existence of clinicocytogenetic syndromes. These patients exhibit specific chromosomal alterations with a well-defined hematological picture and clinical behavior. In this article the clinical implications of the various types of specific chromosomal abnormalities seen in acute nonlymphocytic leukemia, as found in our laboratory and as reported in the literature, are reviewed and discussed.

Acute Disease↗

PubMed1980

Acute unclassified leukemia: A clinicopathologic study with diagnostic implications of electron microscopy.

By rigid cytological and cytochemical criteria, the diagnosis of acute and undifferentiated leukemia was established in 22 patients. According to defined criteria, the leukemic cells could not be classified by conventional light microscopic techniques employed in the study of hematopoietic tissue. Cytochemical studies including peroxidase, periodic acid schiff (PAS) and nonspecific esterase (alpha napthyl butyrate-reacting esterase) stains were done on fresh bone marrow samples, and the percentage of positive leukemia cells for each of these stains was determined on 200 cells. In this series of leukemias, cytochemistry at the light microscope level did not contribute to further classification. Subsequent electron microscopic examination of bone marrow samples from these patients confirmed the immaturity and nuclear/cytoplasmic asynchrony of the leukemic cells. Several in vivo neoplastic markers, such as nuclear blebs, increased nuclear bodies, and cytoplasmic fibrillar bundles could be demonstrated in these cells. Fourteen cases from this series exhibited peroxidase-positive developmental granule formation at the ultrastructural level and were reclassified as acute granulocyte leukemia (AGL). One case was reclassified as lymphoma (poor differentiated type), one case was diagnosed as acute monocytic leukemia (AmonoL), and six cases remained in the undifferentiated category (AUL). Clinical and laboratory features, response to treatment, and survival data were evaluated for these patients. This study demonstrated that electron microscopy is useful in the cytological diagnosis of human leukemia.

Acute Disease↗

PubMed1980

Tumor cell phagocytosis. Its occurrence in a patient with medulloblastoma.

Phagocytosis was observed in tumor cells from a metastatic medulloblastoma to the bone marrow. Erythrocytes and leukocytes were seen in the cytoplasm of the tumor cells. Another abnormal finding was self-phagocytosis, in which tumor cells engulfed one another. The mechanism of tumor phagocytosis is not yet clearly understood. Only a few cases of erythrophagocytosis by epithelial tumor cells have been reported in the literature. To our knowledge, this is the first reported case of a medulloblastoma with hematophagocytosis.

Adult↗