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Biomedical subjects

M L Graham

Publications and source records attributed to M L Graham.

70 records · Page 4Linked to original sources
PubMed1988

Asynchronous antigen expression in B lineage acute lymphoblastic leukemia.

Cell surface phenotypes of 113 B lineage acute lymphocytic leukemia (ALL) cases, defined by the presence of HLA-DR and at least one B-cell-specific antigen (either CD19, CD20, or CD22), were compared with antigen-defined stages of normal B lymphocyte development. The cases were first evaluated for expression of HLA-DR, CD19, CD34, CD10, CD20, and CD22 by indirect one-color immunofluorescence. Pairwise comparisons of cell surface marker expression were performed for each leukemic sample: no correlations were observed for paired antigen expression on the leukemic samples using antigens expressed either early or late during normal B lymphoid development. Complete immunophenotypes of the cases were then compared with normal B-cell developmental stages. Sixteen different complete immunophenotypes were observed on the leukemias that were not found in normal marrow; at least 78% of the cases demonstrated such "asynchronous" combinations of B lymphoid-associated differentiation antigens. Several samples were subsequently studied by two-color immunofluorescence, and the presence of doubly labeled cells with "asynchronous" antigen combinations was confirmed. These results indicate that the majority of B lineage leukemias exhibit "developmental asynchrony," as compared with normal marrow B cells. The data further suggest that ALL cases do not accurately represent cells arrested at the stage where the leukemogenic event occurred. Rather, ALL appears to be a disease in which there may be maturation of leukemic blasts; but this maturation is "asynchronous" when compared with the normal developmental process.

Adult↗
PubMed1988

Microassay for nuclear binding of steroid receptors with use of intact cells from small samples of avian and human tissue.

A micro version of a nuclear binding assay to assess the biological activity of receptors for steroid hormones was developed for application to small (needle) biopsies of human tumors for the purpose of predicting responses to steroid therapy. This easier assay requires 10-fold less tissue than the original nuclear binding assay described for progesterone receptors in the avian oviduct, endometrium, and endometrial carcinomas (Spelsberg TC, et al., Endocrinology 1987;121:631). We describe the application of this micro assay to normal avian oviduct and cancers of the human breast, and we demonstrate a tissue specificity and saturation of nuclear binding. The micro assay reliably measured as little as 0.5 mg equivalents of tissue per assay tube. Results for breast tumors determined to be estrogen-receptor-positive by the standard dextran-coated charcoal method were also determined with this nuclear binding assay. As described previously for progesterone receptors in endometrial carcinomas, some receptor-positive breast biopsies displayed negligible capacity for nuclear binding. Therefore, with the present assay we have identified nonfunctional receptors in these biopsies, which may be useful for accurate prediction of patients' responses to therapy with hormones.

Animals↗
PubMed1987

A nuclear binding assay to assess the biological activity of steroid receptors in isolated animal and human tissues.

This paper describes a nuclear binding assay (NB assay) which measures not only the presence of a steroid receptor in a tissue, but also the quantity of that receptor which is biologically active or functional, i.e. able to bind to nuclear acceptor sites. The assay involves the isolation viable cells from tissues and their incubation with an excess of radiolabeled steroid to encourage the activation and nuclear binding of all cellular receptors. The nuclei are isolated under conditions that remove unactivated (unbound) steroid-receptor complexes. This NB assay demonstrates, in both animal and human steroid target tissues, a saturable, tissue- and steroid-specific, and temperature- and time-dependent nuclear binding of radiolabeled steroids. These properties support a receptor-dependent nuclear binding of steroids. This assay is reproducible and requires relatively small amounts of tissue. The patterns of nuclear binding of the progesterone receptor, achieved with the assay in the avian oviduct model system, are shown to correlate with the nuclear binding of progesterone in vivo, the ability of the steroid to alter transcription, and the expression of a specific gene product, the protein avidin. The assay has been used to identify the existence of nonfunctional steroid receptors in endometrial and breast carcinomas. Therefore, this NB assay combined with the standard charcoal/hydroxylapatite methods of quantitating total cellular receptors should provide a means of assessing changes in the regulation of the biological activity of steroid receptors. Further, the assay should be useful to assess the ability of steroid analogs to properly activate their respective receptors for subsequent nuclear binding.

Animals↗
PubMed1986

Immunoglobulin gene rearrangements in remission bone marrow specimens from patients with acute lymphoblastic leukemia.

Recombinant DNA probes for the joining (JH) segment of the immunoglobulin heavy chain gene were used to detect molecular rearrangements of this gene in the DNA of bone marrow cells obtained during remission of acute lymphoblastic leukemia (ALL). This molecular approach was optimized and found to exceed the sensitivity of conventional morphologic screening for detecting residual leukemia cells; one leukemic cell in 500 normal nucleated bone marrow cells was easily detected using this approach. In the present study, bone marrow from three of seven patients in complete clinical remission (defined morphologically) contained leukemic cells in these proportions. This analysis may be of use in evaluating the status of clinical remission in selected ALL patients.

Adolescent↗
PubMed1986

Isotretinoin and Staphylococcus aureus infection. A possible association.

The use of isotretinoin (13-cis-retinoic acid) in the treatment of numerous dermatologic disorders, as well as the side effects encountered with use of the drug, have increased remarkably since its release. We encountered a case of Staphylococcus aureus endocarditis in a patient with chronic stable aortic insufficiency undergoing therapy with isotretinoin for extensive actinic keratoses. Although significant dysfunction of the immune system has not been demonstrated with isotretinoin, nasal colonization with S aureus has been shown to occur. Changes in skin fragility caused by the drug may provide a portal of entry for the organism. Physicians should be alert for this potential complication in patients with an underlying cardiac valvular lesion; antibiotic prophylaxis may be indicated in this group during isotretinoin therapy.

Aortic Valve Insufficiency↗
PubMed1985

Response of a recurrent choroid plexus tumor to combination chemotherapy.

Choroid Plexus tumors are rare. Surgery and biopsy is diagnostic, and radiotherapy has been used as the treatment of choice for choroid plexus carcinoma (CPC) and recurrent choroid plexus papilloma (CPP). We report the first case of CPP responding to combination chemotherapy consisting of cisplatin, bleomycin and vinblastine (CBV). This chemotherapy regimen should be considered for future trials in patients with choroid plexus tumors and recurrence after surgery and/or radiotherapy.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1985

Blastoderm-differential and blastoderm-specific genes of Drosophila melanogaster.

We have isolated, by molecular cloning, genes expressed differentially at the blastoderm stage of Drosophila melanogaster. Two of the blastoderm-differential genes are reexpressed at later stages, and map to single chromosomal loci 95C and 99E. The sequence at 99E is that encoding the myosin light chain 2. Two other blastoderm-differential sequences are members of multigene families (one of which is B104, or roo) and map to multiple dispersed chromosomal loci. A gastrula-differential sequence was found which maps to 71A. Most significantly, we have identified three genes encoding transcripts expressed uniquely at the blastoderm stage; these map to single chromosomal loci: 25D3, 75C, and 99D4-8. At least some of the blastoderm-differential and blastoderm-specific loci appear to be distinct from loci involved in embryonic pattern formation that have been identified in recent genetic "saturation" screens. The procedure of identifying genes specific to the blastoderm stage may thus allow the identification of genes, not previously identified by classical genetic techniques, that are involved in important embryonic processes.

Animals↗
PubMed1984

Androgen insensitivity syndrome: gonadal androgen receptor activity.

To determine whether abnormalities of the androgen receptor previously observed in skin fibroblasts from patients with androgen insensitivity syndrome also occur in the gonads of affected individuals, androgen receptor activity in the gonads of a patient with testicular feminization syndrome was investigated. Using conditions for optimal recovery of androgen receptor from human testes established by previous studies, we detected the presence of a high-affinity (dissociation constant = 3.2 X 10(-10) mol/L), low-capacity (4.2 X 10(-12) mol/mg DNA), androgen-binding protein when tritium-labeled R1881 was incubated at 4 degrees C with nuclear extracts from the gonads of control patients or from a patient with testicular feminization syndrome but not when incubated at 37 degrees C. Thus this patient has an androgen receptor with a temperature lability similar to that of receptors from normal persons.

Adolescent↗
PubMed1984

Evidence for an androgen receptor in human testis.

The present study identified and characterized an androgen-binding protein in human testicular tissue. Human testes were homogenized in dilute Tris buffer containing thioglycerol, phenylmethylsulfonylfluoride, and molybdate. The supernatant (termed cytosol) was incubated with radiolabeled androgen methyltrienolone (tritium-labeled R1881), and nonspecific binding was determined by adding 100-fold excess of unlabeled R1881 together with [3H]R1881 to cytosol. Specific binding with saturation at 24 hours was observed. Scatchard analysis of the specific binding with the use of increasing concentrations of [3H]R1881 alone and [3H]R1881 plus 200-fold excess unlabeled R1881 demonstrated a high-affinity (dissociation constant = 2.18 X 10(-10) mol/L), low-capacity (2924 molecules per cell) class of binding sites. A second class of lower-affinity sites was identified with a dissociation constant equaling 1.2 X 10(-8) and 26,300 molecules per cell. The bulk of the higher-affinity class of sites was precipitated at 35% ammonium sulfate. In competitive binding assays, dihydrotestosterone and testosterone greatly diminished binding to this high-affinity class of sites. Progesterone also diminished binding but to a lesser degree. Estradiol, estriol, and estrone failed to compete for these sites. Analysis of the receptor, using sucrose gradients, revealed a major peak in the 4S region and a small peak at 8S. A similar high-affinity (dissociation constant = 4.28 X 10(-9), low-capacity (4860 molecules per cell) binding protein was identified in purified nuclei. Binding to nuclear chromatin was demonstrated in the cell-free binding assay, and nuclear binding was further illustrated in the biopsy assay of intact tissue, suggesting translocation in vivo. These properties are characteristic of the androgen receptor and suggest that human testis is a target tissue for androgen, as has been found in animal tissue.

Aged↗
PubMed1984

Transcription, export and turnover of Hsp70 and alpha beta, two Drosophila heat shock genes sharing a 400 nucleotide 5' upstream region.

A highly homologous 400 nucleotide sequence flanks the 5' end and extends 64 NT into the transcribed portion of all five hsp70 and seven alpha beta heat shock genes in Drosophila melanogaster (1-4). To determine the extent to which this sequence dictates coordinate regulation, we compared the total mass, continuous labeling and pulse-labeling of hsp70 and alpha beta RNAs at different times and temperatures of heat shock. By all these measurements, expression of both hsp70 and alpha beta genes increased and decreased in parallel. Hsp70 RNA was generally synthesized at a higher rate and accumulated to a greater extent than alpha beta RNA. As the temperature of heat shock increased, however, the rate of synthesis and accumulation of hsp70 relative to alpha beta RNA decreased. Another difference was that a larger fraction of hsp 70, as compared to alpha beta RNA was exported from the nucleus. For both RNAs, export decreased as the heat shock temperature was increased. The hsp70 and alpha beta genes are thus expressed in parallel, but the homologous 5' upstream sequences do not dictate equal rates of transcription or export from the nucleus.

Animals↗
PubMed1984

Pulmonary lymphangiomyomatosis: with particular reference to steroid-receptor assay studies and pathologic correlation.

The symptoms of progressive dyspnea, hemoptysis, spontaneous pneumothorax, chylous effusions, and cough in conjunction with ventilatory obstruction and abnormal gas exchange in a young woman should prompt the diagnosis of pulmonary lymphangiomyomatosis. Cytosol steroid-receptor assays and postmortem studies were conducted in an extensive investigation of a case of this disease. A biopsy specimen of the lung disclosed evidence of nuclear translocation of [3H]progesterone and the presence of a cytosolic receptor for progesterone, an indication that this disease could be treated effectively with progestin.

Adult↗
PubMed1982

Pharmacology and toxicity of a potent "nonclassical" 2,4-diamino quinazoline folate antagonist, trimetrexate, in normal dogs.

The pharmacology of trimetrexate (JB-11, NSC 249008, 2,4-diamino-5-methyl-5-[(3,4,5-trimethoxyanilino)methyl]quinazoline), an antitumor agent effective against several mouse tumors, was studied in normal dogs. A high-performance liquid chromatographic technique with electrochemical detection, dihydrofolate reductase inhibition assay, and 14C-labeled drug were used to measure plasma disappearance, tissue distribution, excretion, and metabolism of the drug at doses from 0.5 to 6 mg/kg. Doses of 2 mg/kg were well tolerated without toxicity. Higher doses (3 to 6 mg/kg) produced mainly intestinal toxicity without significant hematological or liver abnormalities. The 6-mg/kg dose caused severe bloody diarrhea. After administration of 3 mg/kg, plasma concentrations of trimetrexate were 1 microM and were equal to or greater than 0.1 microM at 1 and 24 hr, respectively. The predominant pharmacokinetics of trimetrexate plasma disappearance was an elimination phase with a t1/2 of 3.5 hr. Concentrations in the cerebrospinal fluid were 2 to 5% of that in plasma and were maximum within 1 to 2 hr after i.v. administration. Highest tissue concentrations of drug were measured in liver and kidney; lowest were found in brain and lung. A dose equivalent to 3 mg/kg in humans (on a sq m basis) should produce adequate plasma concentrations (greater than 0.1 microM) for therapeutic effects.

Administration, Oral↗
PubMed1980

Transcription and metabolism of RNA from the Drosophila melanogaster heat shock puff site 93D.

Characteristics of the major heat shock puff site 93D and the RNA transcribed from it have been investigated by hybridization to polytene chromosome preparations and to recombinant DNA. By saturation in situ hybridization, the length of the transcribed region at 93D is twice that of the mRNA coding region at the heat shock puff site 87A. From the known length of the heat shock mRNA sequence at 87A, we calculate that the minimum length of the transcribed region at 93D is 9.6 kb (kb = kilobase, i.e., 1,000 nucleotides).--The metabolism of RNA transcribed from 93D has been compared with that of RNA coding for the major heat shock protein hsp70 in cells incubated for one hour at 35 degrees C. Hsp70 mRNA sequences, assayed by hybridization to a specific recombinant DNA probe and by in situ hybridization to 87A, were found in both poly(A)+ and poly(A)- cytoplasmic RNA and were more concentrated in cytoplasmic RNA than in nuclear RNA. In contrast, sequences complementary to 93D, assayed by in situ hybridization, were more concentrated in nuclear than in cytoplasmic RNA. This implies that sequences from 93D exit from the nucleus at a lower rate and/or are turned over in the cytoplasm at a higher rate, than sequences from 87A. Site 93D is also unusual in that its transcribed region is represented in both poly(A)+ and poly(A)- nuclear RNA, even though 93D-complementary RNA in the cytoplasm is predominantly poly(A)-. Finally, only 28-58% of the length of DNA transcribed at 93D is represented in cytoplasmic RNA, indicating that only a portion of the sequences transcribed from 93D are exported from the nucleus. The transcripts from two heat shock loci, 93D and 87A, thus appear to be metabolized in significantly different ways.

Animals↗
PubMedDate not supplied

Use of amphotericin B colloidal dispersion in children.

PURPOSE: To describe the experience with a new lipid-based amphotericin product (amphotericin B colloidal dispersion or ABCD) in children with fever and neutropenia who are at high risk for fungal infection. PATIENTS AND METHODS: Forty-nine children with febrile neutropenia were treated in a prospective, randomized trial comparing ABCD with amphotericin B. An additional 70 children with presumed or proven fungal infection were treated with 5 different open-label studies of ABCD. Patients were registered into these studies for reasons of: 1) failure to respond to amphotericin B; 2) development of nephrotoxicity or preexisting renal impairment; or 3) willingness to participate in a dose-escalation study. Extensive data detailing response and toxicity were collected from each patient. RESULTS: In the randomized trial, there was significantly less renal toxicity in the children receiving ABCD than in those receiving amphotericin B (12.0% vs. 52.4% [P = 0.003]). Other adverse symptoms were not significantly different. In the additional open-label studies, although 80% of patients receiving ABCD reported some adverse symptom, the majority of these were infusion related, and nephrotoxicity was reported in only 12% of these patients. CONCLUSIONS: ABCD was well-tolerated at doses up to 5 times greater then those usually tolerated with amphotericin B. Renal toxicity was markedly less than expected, and there were no other unexpected severe toxicities. Further randomized studies are needed to further define the role of this and other liposomal products in children.

Adolescent↗
PubMedDate not supplied

Leukemic iris infiltration.

Three patients with acute lymphoblastic leukemia and leukemic infiltration of the iris are presented. The clinical features, diagnostic techniques, and treatment of this condition are described.

Acute Disease↗