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Biomedical subjects

M L Graham

Publications and source records attributed to M L Graham.

At least 55 records · Page 3Linked to original sources
PubMed1991

Phase II trial of mitoxantrone in acute lymphocytic leukemia of childhood. A Pediatric Oncology Group study.

Thirty children with refractory acute lymphocytic leukemia (ALL) were treated with mitoxantrone, 8 mg/m2/day, for 5 days. Three children received a second course of the drug 3 to 4 weeks later. All but two patients had received prior anthracycline therapy. There were 2 complete responses, 4 early deaths, and 24 patients with persistent leukemia. Of the 21 patients with circulating blasts at the start of mitoxantrone who did not achieve remission, 16 (76%) had complete clearance of their peripheral blood blasts. Although all patients developed profound neutropenia (less than 100 per mm-3), mucosal and hepatic toxicities were uncommon and mild. Mitoxantrone has moderate activity in childhood ALL and should be considered for further trials in less heavily pretreated patients.

Adolescent↗
PubMed1991

A method to analyze 2-dimensional daily radiotherapy portal images from an on-line fiber-optic imaging system.

On-line radiotherapy imaging systems allow convenient treatment verification and generate a wealth of data. Quantitative analysis of data will provide important information about the nature of treatment variations. Using an inhouse fiber-optic imaging system to acquire daily portal images for five patients, we have developed a method to analyze the cumulative positional variation of blocks in the 2-dimensional images. For each beam arrangement used to treat a particular patient, a reference portal image was established. All other images for that patient were registered with respect to the anatomical landmarks visible on the reference image. Two-dimensional frequency distributions describing the overlap of the blocks during the course of treatment were then calculated and superimposed on the reference image. Results of the analysis show positional and quantitative information about the daily variation in block placement, and appeared to be site-dependent. Long term verification studies using on-line imaging systems will be important in the understanding of treatment uncertainties.

Radiology Information Systems↗
PubMed1991

Non-Hodgkin's lymphoma. Biologic classification and implication for therapy.

The non-Hodgkin's lymphomas are a group of diseases for which substantial progress has been made in understanding tumor biology and effectiveness of treatment during the last decade. These advances may provide insight into the development of neoplasms because of recognized association of lymphomas with viral infections and immunodeficiency. The prognosis for patients with NHL continues to improve. As a result, current studies on treatment of lymphomas in certain favorable stages have concentrated on reducing the intensity of therapy. For patients with advanced disease, further improvements in treatment are still being sought.

Bone Marrow Transplantation↗
PubMed1990

T47DCO cells, genetically unstable and containing estrogen receptor mutations, are a model for the progression of breast cancers to hormone resistance.

We postulate that one mechanism for the progression of breast cancers to hormone resistance involves the acquisition of mutant estrogen receptors (ER)4 by genetically unstable cell subpopulations. The T47D human breast cancer cell line may be a model for such progression, having sublines that are ER positive and estrogen responsive, ER positive and estrogen resistant, or ER negative. Also, T47D cells can be either hyperdiploid (HD) or hypertetraploid (HT) or persistently alternate between these states. T47DCO cells are a HD and ER-positive, but estrogen-resistant, subline of T47D cells that undergoes spontaneous tetraploidization. Such a stable variant, designated T47Dv, is 85% HT (Cancer Res., 49: 3943, 1989). We now show that single-cell clones derived from the mixed HD/HT T47Dv can be either HD or HT, and can be either estrogen responsive or estrogen resistant, for growth and for progesterone receptor regulation. To begin the study of ER in this model system of T47DCO and their derivatives, we have generated complementary DNA libraries from the parental HD T47DCO cells and have isolated three ER complementary DNA mutants. These include two frame-shift/termination mutants that would encode ERs truncated in the DNA-binding domain and in the hormone-binding domain and a third mutant with a large in-frame deletion spanning the hinge region and a part of the hormone-binding domain. If expressed, these mutant ERs would lack hormone-binding capacity and would be undetected by the anti-ER antibodies currently in clinical use. Genetic instability, when associated with mutant ERs in subpopulations of breast tumor cells, may provide the selective pressure leading to hormone resistance. T47DCO cells and their clonal derivatives provide a model for the systematic study of ER mutations and other mechanisms of hormone resistance in Stage IV breast cancer.

Amino Acid Sequence↗
PubMed1990

Identification of putative nonfunctional steroid receptors in breast and endometrial cancer.

A nuclear binding assay was developed for the purpose of having available a more predictive assay for hormone responsiveness in human cancers. The BNB assay identified specific and saturable steroid nuclear binding in human target tissues and human carcinomas. When the BNB assay was applied to a large set of breast and endometrial carcinomas, we speculate that nonfunctional receptors were detected in 20%-50% of the patients who were receptor-positive by the DCC assay. Lastly, as responsiveness to hormonal therapy in these cancer patients becomes known, the predictive value of the BNB assay can be established.

Biopsy↗
PubMed1990

Invasive fungal disease in pediatric acute leukemia patients with fever and neutropenia during induction chemotherapy: a multivariate analysis of risk factors.

We evaluated the courses of 115 consecutive cases of pediatric acute leukemia treated with induction chemotherapy. Seventy-two patients developed fever associated with neutropenia; 15 developed systemic fungal infections. We reviewed multiple demographic and treatment characteristics of these patients in an attempt to identify potential risk factors for the development of invasive fungal disease (IFD). Risk factors identified in a univariate analysis included duration of neutropenia after first fever (P less than .0001), diagnosis of acute nonlymphocytic leukemia (ANLL) (P = .003), onset of fever and neutropenia within 5 days of starting induction chemotherapy (P = .009), and multiple (greater than one) surveillance culture sites positive for fungal organisms (P = .02). In a multiple logistic regression analysis, duration of neutropenia (P less than .001) remained a significant risk factor. The study group of patients had a significantly higher risk of fungal infections than a matched group of leukemia patients developing fever with neutropenia due to postremission consolidation chemotherapy (P = .003). In the first 48 patients, 14 (29%) developed IFD. In the subsequent patients (n = 24), intravenous miconazole (5 mg/kg every 8 hours) was begun at the time of the first fever. One of the 24 patients (4%) given miconazole developed IFD. The use of miconazole was a negative risk factor for the development of IFD in univariate (P = .01) and multivariate (P = .05) analysis. We conclude that pediatric leukemia patients who develop fever associated with neutropenia during induction chemotherapy are at high risk for developing IFD. The role of intravenous miconazole at the time of the first fever in this group deserves further study.

Adolescent↗
PubMed1989

Simultaneous measurement of progesterone receptors and DNA indices by flow cytometry: analysis of breast cancer cell mixtures and genetic instability of the T47D line.

A flow cytometry assay that can simultaneously measure progesterone receptors (PR) and DNA indices in breast cancers would be a valuable clinical tool. We have developed a prototype assay that has proven useful in studies of the cell biology of breast cancer cell lines. The assay uses PR-specific monoclonal primary antibodies and fluorescein-conjugated secondary antibodies to measure PR, and propidium iodide to measure DNA. We find that the specific PR fluorescence generated by labeling PR-rich T47D human breast cancer cells is located predominantly in nuclei. The flow cytometry assay can quantitatively measure large fluctuations in intracellular PR levels: an apparent increase in PR following acute progestin treatment that cannot be documented by ligand binding assays; and the receptor down-regulation that follows chronic progestin treatment. The assay can identify fewer than 10% PR-positive cells in a population of PR-negative cells having the same DNA content, and it can sort PR-positive and PR-negative cells from cell mixtures having different DNA indices. Gating allows quantitative analysis of these mixed cell populations by ploidy, cell-cycle phase, and PR content. Finally, the assay has allowed us to monitor the gradual emergence of a stable hypertetraploid cell population, designated T47Dv, from the wild-type hyperdiploid T47Dco stocks. The new cells have unchanged estrogen receptors but even higher PR levels than the parental cells. They have five to ten copies of chromosome 11, site of the PR gene and other genes of interest in breast cancer.

Biomarkers, Tumor↗
PubMed1989

Simultaneous measurement of progesterone receptors and DNA indices by flow cytometry: characterization of an assay in breast cancer cell lines.

Progesterone receptors (PR) are the strongest predictors of response to hormone therapy in metastatic breast cancer, while PR and the DNA indices of cell ploidy and percentage of S phase are useful prognostic indicators in early stage breast cancer. We have developed a flow cytometry method to measure PR and DNA indices simultaneously using two aneuploid breast cancer cell lines--PR-positive T47D cells and PR-negative MDA-231 cells. Cells were pretreated with the progestin 17,21-dimethyl-19-nor-4,9-pregnadiene-3,20-dione, harvested, counted, fixed with paraformaldehyde, and permeabilized with Triton X-100. To measure total PR, cells were first exposed to a mixture of the mouse anti-PR monoclonal antibody AB-52, which binds both Protein A and Protein B of human PR, and to monoclonal antibody B-30 or B-64, which bind only Protein B. Then the cells were treated with fluorescein isothiocyanate-conjugated goat anti-mouse second antibody to produce a green fluorescence signal corresponding to PR. To measure nonspecific binding, cells were treated with mouse IgG1 as the first antibody in a parallel incubation. Specific immunoassayable PR is the difference between total and nonspecific binding. Following the antibodies, the cells were treated with RNase A and propidium iodide to give a red fluorescence signal corresponding to DNA content. Red and green fluorescence per cell was then quantified by flow cytometry. This method gives a strong specific signal for PR in several T47D cell sublines but no specific binding in MDA-231 cells. Progestin treatment led to apparent increases in PR. The proportion of cells in the G0-G1, S, and G2-M phases of the cell cycle was determined from DNA histograms and showed that both cell lines were hyperdiploid. The simultaneous flow cytometry method allowed assignment of relative PR levels in subsets of cells segregated by their DNA content. In T47D cells, PR were present throughout the cell cycle, and levels doubled in G2 and mitosis.

Antibodies, Monoclonal↗
PubMed1989

Phase I trial of trimetrexate glucuronate on a five-day bolus schedule: clinical pharmacology and pharmacodynamics.

Trimetrexate glucuronate (TMTX), a nonclassical folate antagonist, has been evaluated clinically on several schedules. We have studied TMTX administered as an iv bolus for 5 consecutive days every 3 weeks in 35 patients with advanced solid tumors. Drug was given at doses ranging from 7.6 to 18.8 mg/m2. The maximal tolerated dose was 13.1 mg/m2 per day x 5 for patients without prior myelotoxic treatment and 7.6 mg/m2 per day x 5 for previously treated patients. Because of wide individual differences in drug tolerance, dose escalation in 25% increments is recommended for patients not experiencing toxic effects. The dose-limiting toxicity was neutropenia. Rash and mucositis were also significant. TMTX concentrations were measured 1 and 24 hours after each dose, and the data were fit by use of a one-compartment pharmacokinetic model. With this simplified sampling and modeling scheme, the mean total body clearance (+/- SD) of trimetrexate was 31 +/- 20 mL/min per m2 and the volume of distribution was 13 +/- 7 L/m2. Mean plasma concentrations 1 hour after a dose were 1.12, 2.43, 3.33, and 3.25 mumol/L at 7.6, 9.1, 10.9, and 13.1 mg/m2, respectively. The mean TMTX concentration (+/- SD) 24 hours after a dose was 114 +/- 87 nmol/L. The mean area under the concentration-versus-time curve at 13.1 mg/m2 was 2,266 mumol.min/L. The drug concentration 1 hour after the first dose and the area under the concentration-versus-time curve were highly correlated with leukopenia and thrombocytopenia (r = .6 and .65 and P = .0007 and .0001, respectively). The maximal tolerated dose on the daily x 5 schedule was 30% of the dose tolerated on an iv bolus schedule. The choice of drug schedule for clinical trials when murine and human pharmacokinetics differ is discussed. Phase II trials are under way with both the iv bolus and the daily x 5 schedules.

Adult↗
PubMed1989

Ophthalmic manifestations of leukemia.

We prospectively examined the eyes of 120 patients with leukemia. Only 38% of patients had normal results of examination. Leukemic infiltrates were present in 3% of patients, other findings related to leukemia were seen in 39% of patients, and 20% of patients had unrelated abnormalities. Visual loss was seen in at least 5% of the patients in this study.

Adolescent↗
PubMed1989

Intraocular penetration of cytosine arabinoside following subconjunctival administration in primates.

Cytosine arabinoside was administered subconjunctivally to three monkeys. Blood, aqueous, and vitreous specimens were collected after 30 minutes, 1 hour, 2 hours, and 3 hours. Intraocular drug concentrations appeared stable or increasing over this time period while intravenous levels were relatively low. The results of this study suggest that therapeutic intraocular concentrations of cytosine arabinoside are obtained following subconjunctival administration in primates with lower, nontoxic intravenous levels during this time.

Animals↗
PubMed1989

Estrogen and progesterone receptors in meningiomas: comparison of nuclear binding, dextran-coated charcoal, and immunoperoxidase staining assays.

We studied the status of estrogen (ER) and progesterone (PR) receptors in meningiomas removed from 52 patients, comparing dextran-coated charcoal (DCC), nuclear binding (NB), and immunoperoxidase (IP) assays. Each of the assays was performed independently by investigators well-experienced with these assays. The NB assay is a new assay that measures functional steroid receptors--that is, the activation of the receptor and its binding to the nucleus. The assay is very sensitive and requires a relatively small amount of tissue as compared with the DCC assay. In agreement with data from other studies. PR were detected in most meningiomas by all 3 methods: in 69% of the cases by NB, in 76% by DCC, and in 89% by IP. ER were detected in only a few cases: in 33% by NB, in 2% by DCC, and in none by the IP assay. The agreement for PR sites was 62% for all 3 assays; it was 66% between the NB and DCC assays, 67% between the NB and IP assays, and 86% between the DCC and IP assays. Of 26 cases that were positive by the DCC assay, 6 (23%) were negative by NB. The overall agreement for all three ER assays was 65%. The data suggest that the majority of meningiomas contain high-affinity receptors for progesterone, that estrogen receptors are present in only a few meningiomas, and that some of these estrogen and progesterone receptors appear to be functional.

Adolescent↗
PubMed1988

Evidence of estrogen receptors in normal human osteoblast-like cells.

In seven strains of cultured normal human osteoblast-like cells, a mean of 1615 molecules of tritium-labeled 17 beta-estradiol per cell nucleus could be bound to specific nuclear sites. The nuclear binding of the labeled steroid was temperature-dependent, steroid-specific, saturable, and cell type-specific. These are characteristics of biologically active estrogen receptors. Pretreatment with 10 nanomolar estradiol in vitro increased the specific nuclear binding of progesterone in four of six cell strains, indicating an induction of functional progesterone receptors. RNA blot analysis demonstrated the presence of messenger RNA for the human estrogen receptor. The data suggest that estrogen acts directly on human bone cells through a classical estrogen receptor-mediated mechanism.

Binding, Competitive↗
PubMed1988

A nuclear binding assay for measurement of biologically active androgen receptors in animal tissues and human prostate cancer.

A nuclear binding (NB) assay has been developed for the measurement in intact viable cells of biologically active (functional) estrogen and progesterone receptors, i.e. those capable of binding to nuclear acceptor sites [Spelsberg et al., Endocrinology 121: 631 (1987)]. This paper describes the application of this assay to analyses of androgen receptors in the guinea pig seminal vesicle and in human prostatic carcinoma. Cells from fresh animal seminal vesicles or human prostate carcinoma are isolated using collagenase and are incubated with [3H]R1881 for 1 h at 22 degrees C, after which nuclei are isolated at 4 degrees C and assayed for DNA and radioactivity. This NB assay demonstrates a saturable, temperature dependent, steroid and tissue specific nuclear binding of [3H]R1881 for the guinea pig-seminal vesicle system. The nuclear binding is of high affinity and low capacity. The NB assay reveals several important aspects of the androgen and estrogen receptors in target tissues: (1) the nuclear acceptor sites for androgen receptor (AR) are steroid receptor specific; (2) there are different concentrations of the androgen and estrogen receptors between the epithelium and the fibromuscular components of the guinea pig seminal vesicle; and finally (3) some biopsies of human prostate cancer appear to contain biologically inactive AR. This assay may be useful in the analyses of functional receptors in biopsies of human cancer cells.

Animals↗
PubMed1988

Intraocular penetration of cytosine arabinoside following intravenous administration in primates.

Cytosine arabinoside was administered intravenously to three monkeys. Blood, aqueous, and vitreous specimens were collected after 30 minutes, 1 hour, 2 hours, and 3 hours. Intraocular and intravenous drug concentrations were comparable at all time points, at levels likely to be effective against human tumors. This study suggests that therapeutic intraocular concentrations of cytosine arabinoside are attained following intravenous administration in primates.

Animals↗
PubMed1988

Acute leukemia expressing the normal human hematopoietic stem cell membrane glycoprotein CD34 (MY10).

Thirty-one cases of acute leukemia with blast cells greater than or equal to 70% positive for the hematopoietic stem cell Ag, CD34 (MY10, HPCA-1), were identified from the University of Nebraska Medical Center and The Johns Hopkins Oncology Center over an 18-month period. Fourteen of the cases were classified as early B-lineage ALL, 3 cases were other ALL subtypes, and 14 of the cases were ANLL. Five of the 17 cases of ALL expressed one or more myeloid-associated surface Ags, 3 ANLL cases expressed CD10 (CALLA, J5), and T-lymphoid Ags were present in 12 of 31 cases (1 T-cell ALL, 3 of 16 B-lineage ALL cases, and 8 of 14 ANLL cases). Eleven of 12 CD34+ ALL cases studied had abnormal karyotypes; only 7 of 12 CD34+ ANLL cases studied had abnormal karyotypes, and 3 of these were CD10+ ANLL. Six cases were Ph1 positive, including the one mature B cell ALL, 4 early B-lineage ALL, and 1 CD10+ ANLL case. Good and poor prognosis subgroups of high frequency of expression of CD34 leukemias could be identified, generally, as would have been predicted by previously defined criteria. Thus, of the 10 Ph1-negative early B-lineage ALL patients, 9 achieved CR (90%). At the other extreme, the CR rate of CD10- ANLL was 4 of 11 (36%). The leukemias characterized by greater than or equal to 70% of cells positive for CD34 form a relatively undifferentiated subset of the leukemias which may show features associated with more than one lineage, and if CD10- and myeloid morphology, may respond poorly to therapy.

Antigens, CD34↗
PubMed1988

Cytomegalovirus infection after autologous bone marrow transplantation with comparison to infection after allogeneic bone marrow transplantation.

Cytomegalovirus (CMV) infection was detected in 65 of 143 (45%) autologous bone marrow transplant (BMT) patients. CMV pneumonitis occurred in only 2% of the patients and CMV retinitis occurred in none. Infection occurred in half of the 40 initially seronegative patients and 47% of the 94 initially seropositive patients. Among initially seropositive patients, platelet recovery was slower in infected patients than in those not infected (97 v 35 days median, P = .003), and neutrophil recovery was slightly delayed in infected patients (31 days v 24 days, P = .02). Although the incidence of CMV infection was comparable in autologous and allogeneic BMT patients, CMV pneumonitis was less frequent in autologous BMT patients (2% v 12%, P less than .001). The risk for CMV pneumonitis in autologous BMT patients was comparable with that in allogeneic BMT patients without graft-v-host disease (GVHD) (2% v 6%), but significantly lower than the risk in allogeneic BMT patients with GVHD (2% v 23%, P less than .001).

Bone Marrow Transplantation↗