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Biomedical subjects

R Maki

Publications and source records attributed to R Maki.

35 records · Page 2Linked to original sources
PubMed1993

Regulation of lymphoid-specific immunoglobulin mu heavy chain gene enhancer by ETS-domain proteins.

The enhancer for the immunoglobulin mu heavy chain gene (IgH) activates a heterologous gene at the pre-B cell stage of B lymphocyte differentiation. A lymphoid-specific element, microB, is necessary for enhancer function in pre-B cells. A microB binding protein is encoded by the PU.1/Spi-1 proto-oncogene. Another sequence element, microA, was identified in the mu enhancer that binds the product of the ets-1 proto-oncogene. The microA motif was required for microB-dependent enhancer activity, which suggests that a minimal B cell-specific enhancer is composed of both the PU.1 and Ets-1 binding sites. Co-expression of both PU.1 and Ets-1 in nonlymphoid cells trans-activated reporter plasmids that contained the minimal mu enhancer. These results implicate two members of the Ets family in the activation of IgH gene expression.

Animals↗
PubMed1992

Transection of the stria terminalis without damage to the medial amygdala does not alter behavioural sodium regulation in rats.

Damage to the medial region of the amygdala has been shown to impair mineralocorticoid-induced sodium appetite, while leaving intact sodium appetite induced through sodium depletion. This effect may result from the interruption of the flow of information through the stria terminalis (ST), a neural pathway linking the medial amygdala with the ventral forebrain. We determined the effect of transecting the ST of the rat, at a point remote from the medial amygdala, on sodium appetite induced with the administration of mineralocorticoids and with the natriuretic furosemide. Similar to control and amygdala lesioned rats, rats with ST knife-cuts displayed a normal sodium appetite following treatment with furosemide. However, unlike medial amygdala lesions, transection of the ST alone did not block mineralocorticoid-induced sodium appetite. Therefore, the inability of mineralocorticoids to induce a salt appetite in medial amygdala lesioned rats does not result from damage to the stria terminalis.

Amygdala↗
PubMed1990

Quantitative analysis of cytokine gene expression in rheumatoid arthritis.

Previous studies of the cytokine profile of rheumatoid arthritis (RA) have been primarily limited to the assessment of the levels of these mediators in synovial fluid (SF) or synovial tissues (ST) by biologic or immunologic assays. We have studied cytokine gene expression in RA by in situ hybridization of SF cells, enzymatically dispersed ST cells, and frozen sections of ST. RA ST cells (n = 7) were studied and a high percentage of cells hybridized to the following anti-sense probes: IL-6 = 19 +/- 3.3%; IL-1 beta = 9.9 +/- 1.7%; TNF-alpha = 5.8 +/- 1.4%; granulocyte-macrophage-CSF = 2.2 +/- 0.8%; transforming growth factor-beta 1 = 1.3 +/- 0.2% (p less than 0.05 for each compared to sense probes). Similar results were found using osteoarthritis ST cells, although the percentage of cells expressing the IL-6 gene (7.1 +/- 2.5%) was significantly less in osteoarthritis compared to RA. RA ST cells did not significantly bind the IFN-gamma probe (0.2 +/- 0.1% positive), although they were capable of expressing the IFN-gamma gene if stimulated with PHA. The OKM1+ population of ST cells (i.e., macrophage lineage cells) was greatly enriched for IL-1 beta and TNF-alpha, whereas the OKM1- population (lymphocytes, fibroblasts, and type B synoviocytes) was enriched for IL-6. The vast majority of cells expressing the IL-6 gene were non-T cells. Furthermore, hybridization to RA ST frozen sections localized IL-6 mRNA to the synovial lining layer, which is comprised of type A and type B synoviocytes. In contrast to the high level of cytokine gene expression observed in ST, SF cells did not hybridize significantly to any of the cytokine probes. If stimulated with LPS or PHA, SF cells expressed IL-1 beta or IFN-gamma genes, respectively.

Antigens, Surface↗
PubMed1989

Inhibition of MHC class II-restricted T cell response by Lyt-2 alloantigen.

T cell hybridomas were established by fusing a CD8+ V beta 8.1+ CTL clone and a CD4+ V beta 8.1+ helper T lymphocyte (HTL) clone to the thymoma cell line BW5147. In contrast to the HTL x BW hybridomas, which retain the same antigen specificity as the original T cell clone, the CTL x BW hybridomas lost the class I MHC-restricted antigen response but acquired a new specificity to Mlsa antigen. Mlsa reactivity of CTL x BW hybridomas was shown to be mediated by the CTL TCR as assayed by inhibition using an anticlonotypic antibody to the CTL clone. Since hybridomas established with BW5147 lose CD8 expression, we have introduced the CD8 molecule into CTL x BW5147 hybridomas by gene transfection. The CD8+ V beta 8.1+ hybridoma was no longer capable of reacting to Mlsa antigen but exhibited the same antigen specificity as the parental CTL clone. Furthermore, the presence of the transfected CD8 molecule in the HTL x BW hybridomas was found to be inhibitory to class II MHC-restricted antigen reactivity. These results demonstrate that, besides its role in increasing the overall avidity of T cell-class I MHC/antigen interaction, the CD8 molecule inhibits T cell-class II MHC gene product/antigen interaction. This negative effect of the CD8 molecule on a class II MHC-restricted response may account for the failure of CD8+ T cells using either V beta 8.1 or V beta 6, which impart reactivity to the Mlsa antigen on CD4+ T cells, to respond to the Mlsa antigen.

Animals↗
PubMed1989

Altered I-A protein-mediated transmembrane signaling in B cells that express truncated I-Ak protein.

Recent evidence suggests that the major histocompatibility complex class II molecules of B lymphocytes function as signal-transducing receptors during the generation of T lymphocyte-dependent humoral immune responses. By analogy with other receptors, we postulate that perturbation of the class II molecules is coupled to the generation of intracellular second messengers through interactions involving the transmembrane and/or cytoplasmic domains of the class II molecules. We report a series of experiments that assess which amino acids of the class II molecule I-Ak are required for coupling it to the signal-transduction pathway. We prepared a series of B-lymphocyte transfectants that express I-Ak molecules with COOH-terminal truncations of either the Ak alpha or Ak beta chain or both. The ability of each transfected class II molecule to transduce a signal after being bound by monoclonal antibody was found by monitoring the translocation of protein kinase C from the cytosol to the "nuclear compartment" of the transfected B lymphocyte. Results indicate that the Ak beta chain plays the dominant role in signal transduction and that the 6 cytoplasmic amino acids of Ak beta chain most proximal to the inner plasma membrane are of greatest importance in coupling I-Ak molecules to the molecules of the signaling cascade.

Amino Acid Sequence↗
PubMed1988

Picture memory: recognizing added and deleted details.

When people are presented simple and complex pictures and then tested in a same-changed recognition test with a simple or complex form of each, d' is greater for the simple than the complex picture (Pezdek & Chen, 1982). The results of three experiments confirm the robustness of this "asymmetric confusability effect" and test a model of the processes underlying this effect. According to the model, pictures are schematically encoded such that the memory representation of both simple and complex pictures is similar to the simple form of each. In Experiment 1, a sentence was presented that described the central schema in the picture prior to subjects' viewing each picture. This manipulation exaggerated the asymmetric confusability effect; schematic processing thus underlies the effect. Results of Experiment 2 refute the hypothesis that the effect results from subjects erroneously anticipating a recall test rather than a recognition test. Furthermore, although some of the nonschematic elaborative information in complex pictures is stored in memory, it is difficult to retrieve to verify that something is missing when complex presentation pictures are changed to simple test pictures (Experiment 3). Thus, although people are able to distinguish large sets of old pictures from new distractor pictures, their ability to detect missing elaborative visual details is more limited.

Attention↗
PubMed1987

Comparative sequence analysis of cDNA clones encoding I-A molecules of the CH12 B cell lymphoma: nucleotide differences do not account for their "defective" function in B cell stimulation.

The CH12 B cell lymphoma can be stimulated to secrete antibody by helper T cells that interact with I-EK but not I-AK molecules expressed on its membrane. Both molecules present antigen to the appropriate T cells. We have analyzed the mRNA by Northern blot analysis and obtained partial sequences of cDNA clones encoding A alpha and A beta of the I-A molecule to determine if deletions or mutations in the cytoplasmic or transmembrane domains account for the "defect" in triggering following interaction with I-A restricted helper T cells. The results provide no evidence for structural alterations in either A alpha or A beta which could account for these observations. The implications of these findings on the role of class II molecules in B cell activation is discussed.

Amino Acid Sequence↗
PubMed1981

The role of DNA rearrangement and alternative RNA processing in the expression of immunoglobulin delta genes.

We have established the exon-intron structure of the gene coding for the constant (C) region of the mouse immunoglobulin delta heavy chain, using DNA clones isolated from BALB/c embryos and the delta mRNA extracted from two delta-producing hybridomas, B1-8. delta 1 and GCL2.8. At least three types of C delta gene structures are identified. A 2.7 kb delta mRNA reveals six exons. This delta mRNA may code for a membrane-bound delta chain. A second delta mRNA of 1.8 kb shares the first (5' side relative to direction of transcription) three exons with the 2.7 kb delta mRNA and in addition contains a fourth exon unique to this mRNA species. This delta mRNA most likely codes for a secreted delta chain. A third delta mRNA, also of 1.8 kb, shares the first four exons and a part of the fifth exon with the 2.7 kb mRNA. Its function, if any, remains unclear. We investigated the question of how a lymphocyte can produce the mu and delta heavy chains simultaneously, using the hybridoma GCL 2.8, which makes both IgM and IgD. Results of Southern gel blot analysis and gene cloning experiments indicate that this cell utilizes the same rearranged VH gene for the synthesis of the mu and delta chains, and yet maintains the embryonic configuration for the C mu and C delta genes and for the intervening region. Based on these results, we conclude that the VH sequence is spliced alternatively to the C mu or C delta sequence during processing of the primary RNA transcript. An alternative mechanism for the expression of the delta gene is found in hybridoma B1-8. delta 1, which actively secretes delta chains and synthesizes no mu chain. This mechanism involves deletion of the C mu gene, which brings the complete VH gene closer to the C delta gene.

Animals↗
PubMed1981

Linkage of the four gamma subclass heavy chain genes.

The genes for the heavy-chain constant regions of the four gamma subclass immunoglobulins were identified in a set of overlapping mouse DNA fragments representing about 100 kilobase pairs (kb) of the mouse genome that was cloned from bacteriophage lambda libraries of BALB/c mouse embryo DNA. R-loop mapping studies show that the genes are located 5'-C gamma 3-34 kb-C gamma 1-21 kb-C gamma 2b-15 kb-C gamma 2a-3' and lie in the same transcriptional orientation. Two DNA segments, one of 19 kb and another of 15 kb, that surround the C gamma 2b and C gamma 2a genes, respectively, show considerable homology and implicate a tandem duplication mechanism in the evolution of this gene cluster.

Animals↗
PubMed1980

Immunoglobulin gene rearrangement in immature B cells.

Two types of immature B cells, namely fetal liver hybridomas and the leukemic cell line 70Z/3, both of which have cytoplasmic mu chains but no light chains, were examined for DNA rearrangements of their light chain and heavy chain immunoglobulin genes. In the fetal liver hybridomas, which were constructed from fetal liver cells and a tumor cell, no light chain gene rearrangement was observed, whereas in the 70Z/3 cell line a kappa light chain rearrangement probably occurred. The results suggest that, although the lack of light chain synthesis can be due to a lack of gene rearrangement, there may also be transcriptional regulation, which may also be important for the expression of light chain immunoglobulins in immature B cells.

Animals↗
PubMed1980

Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes.

At least two types of somatic recombination are necessary for the generation of a complete immunoglobulin gamma 2b gene from germ-line DNA sequences. The first type of recombination consists of the assembly of three separate DNA segments, each encoding a different part of the variable region. The second type of recombination replaces the exons coding for the constant region of the mu chain with those coding for the same region of the gamma 2b chain. The DNA sequencing studies suggest that the two types of recombination operate by different mechanisms.

Animals↗
PubMed1980

Exon shuffling generates an immunoglobulin heavy chain gene.

From endonuclease EcoRI partial libraries of DNAs from mouse embryo and MOPC 141, a gamma 2b-producing myeloma, clones were isolated by using a DNA fragment carrying the gamma 2b constant (C) region gene as a hybridization probe. One clone from MOPC 141 contained a heavy chain variable (V) gene and the C gamma 2b gene, as demonstrated by R-loop mapping. The V gene and C gene in this clone were separated by a 3.9-kilobase intron. The characterization of this clone as well as the embryonic clones suggest that at least two recombination events occurred to create the gamma 2b gene in MOPC 141. One of the events is analogous to the V-J joining previously demonstrated in the light chain genes, which brings the major part of the V gene next to a short coding sequence (J). The other event we refer to as "C mu-C gamma 2b switch recombination" because a portion of the intron between the V gene and C gene of the rearranged gamma 2b gene is derived from the 5' flanking sequence of the embryonic C mu gene. A model suggesting how the phenomenon of switch seen in lymphocytes may occur is presented.

Animals↗
PubMedDate not supplied

How a multidisciplinary team improved a process to attain positive patient outcome.

A case study approach will be used to demonstrate the effectiveness of a continuous quality improvement team designed to explore the process of preoperative medical record completeness. The goal of this team was to improve essential chart components (e.g., electrocardiography, history and physical examination) required 48 hours before the date of surgery in an effort to facilitate optimum patient management and timely flow of patients to the operating room. When the 48-hour compliance rule is followed, the nurse reviewer has sufficient time to review all patient data and notify attending physicians or designees of any abnormalities. In essence, improving component compliance provides better quality care because patients are able to be assessed before surgery with enough time to solve any problems.

Aged↗