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Biomedical subjects

J Turner

Publications and source records attributed to J Turner.

At least 181 records · Page 10Linked to original sources
PubMed1996

Evolution of insulin-like growth factor-1, prostaglandin E2, and mitogenic activity of bovine mammary primary lymph during the dry period and lactogenesis.

Four pregnant cows near the end of lactation were fitted with a catheter in a lymph duct afferent to the supramammary lymph node. Cows were dried off 3 d after surgery, and samples of lymph were collected daily from the day of surgery until 4 d postpartum. Samples of blood and mammary secretions were taken before and after drying off and at parturition. Concentrations of most metabolites were lower in lymph than in serum. Concentrations of IGF-I and prostaglandin E2 were not affected at drying off but decreased and increased, respectively, at parturition. All IGF-binding proteins that were present in serum were also present in lymph fluid, but the binding activity was lower. Mitogenic activities of lymph samples taken at various physiological stages were determined on mammary epithelial (MAC-T) and fibroblast cell lines. Lymph was mitogenic, but mitogenic activity was not related to physiological stages. The correlation was high between mitogenic activity of lymph on MAC-T cells and the content of prostaglandin E2 in lymph. Supplementation of lymph with additional prostaglandin E2 increased mitogenic activity, and neutralization of lymph by antibodies reduced mitogenic activity. Basal medium conditioned by the epithelial cell line contained 100 to 250 pg/ml of immunoassayable prostaglandin E2.

Animals↗
PubMed1996

Elective bedside tracheostomy in the intensive care unit.

BACKGROUND: Tracheostomy is a frequently performed procedure, and historically has had a high reported complication rate. This has led some authors to suggest that a tracheostomy should be done only in the operating room (OR). Concerns regarding the hazards of transporting critically ill patients to the OR may inhibit the use of tracheostomy. Bedside tracheostomy in the Intensive Care Unit (ICU) has been shown to be safe, but this concept has not been widely accepted. STUDY DESIGN: We retrospectively reviewed consecutive patients undergoing tracheostomy over a four-year period and compared the safety of elective beside tracheostomy with OR tracheostomy. RESULTS: We studied 536 patients who underwent tracheostomy during the four years, in 470 of whom the procedures were elective. Of these, 66 percent were done at the bedside without an anesthesiologist present. For the 311 patients who underwent bedside tracheostomy, the complication rate was 8.7 percent compared to 9.4 percent of 159 patients undergoing OR tracheostomy (p values were not significant). No deaths were directly associated with tracheostomy procedures, although the overall hospital mortality rate for these patients was 59 percent. CONCLUSIONS: A tracheostomy can safely be performed in an ICU without requiring transport of the patient to the operating suite, thereby eliminating the hazards of transport.

Adolescent↗
PubMed1996

Selected strategies to augment polynucleotide immunization.

We sought to amplify the immune response to polynucleotide immunization through co-delivery of complementary DNA (cDNA) encoding a cytokine or co-stimulatory molecule to enhance antigen presentation. In the context of intramuscular immunization, we examined co-delivery of cDNAs for B7-1 and human carcinoembryonic antigen (CEA) within separate plasmids or a dual plasmid with two independent expression cassettes. Intramuscular delivery of the dual expression plasmid produced anti-CEA antibody responses and antitumor effects superior to those generated by plasmid DNA encoding CEA alone. However, co-delivery of cDNAs encoding B7-1 and CEA in the form of two separate plasmids produced no augmentation. The importance of single plasmid delivery suggests the effectiveness of this strategy is contingent upon co-expression of B7-1 and CEA within the same cell. The success of cutaneous polynucleotide immunization by particle bombardment is thought to derive largely from the presence of Langerhans cells within the skin. We hypothesized that co-delivery of plasmid DNA encoding granulocyte-macrophage colony stimulating factor (GM-CSF) by particle bombardment would enhance the antigen presenting capacity of Langerhans cells at the inoculation site similar to its effects in vitro. Augmentation of CEA-specific lymphoblastic transformation and antibody response was observed when plasmid GM-CSF (pGM-CSF) was administered 3 days prior to each dose of plasmid DNA encoding CEA. These strategies for augmentation of immune response to polynucleotide immunization should be applicable to a wide variety of antigenic targets including infectious agents and other tumor-associated antigens.

Animals↗
PubMed1996

Pulpal response to dentin etched with 10% phosphoric acid.

PURPOSE: To histopathologically evaluate the effect of etching sound dentin with 10% phosphoric acid, in cavities prepared just within the DEJ on young patients. MATERIALS AND METHODS: Twenty-four orthodontic patients who required premolar extractions were selected for the study. Each of the patients had facial Class V resin composite restorations placed in two maxillary premolars selected for extraction. Each preparation was cut circular, 2 mm in circumference and 0.5 mm into dentin. The dentin of one maxillary premolar preparation, randomly selected, was etched with 10% phosphoric acid (Bisco) for 20 seconds. This was followed by the placement of a dentin bonding agent (All-Bond 2) and a microfilled composite restoration (Silux). The contralateral maxillary premolar, received a similar preparation, but the dentin surface was lined with a light-cured glass ionomer liner before etching of the enamel, followed by placement of a dentin bonding agent and a microfilled resin composite restoration. One of the mandibular premolars was randomly selected as a negative control, having no treatment. Fourteen days after the placement of the restorations, the teeth were extracted, and histologically examined for insult to the pulpal tissues. Injury to the pulp was evaluated by quantitative measures of histopathology, including necrosis, infiltration by inflammatory cells, and other indicators of tissue injury. RESULTS: Using a Kruskal-Wallis analysis, there was no statistically significant difference in pulpal histopathology of the teeth with etched dentin versus teeth with dentin protected by a liner or the negative control teeth (P < 0.05).

Acid Etching, Dental↗
PubMed1995

Granulocyte-macrophage colony-stimulating factor mRNA stabilization enhances transgenic expression in normal cells and tissues.

To increase transgenic production of granulocyte-macrophage colony-stimulating factor (GM-CSF), we mutated the mRNA's 3'-untranslated region, AUUUA instability elements. Expression vectors containing human or murine GM-CSF cDNAs coding for wild-type (GM-AUUUA) or mutant versions with reiterated AUGUA repeats (GM-AUGUA) were transfected into cells in culture or animals using particle-mediated gene-transfer technology. Normal peripheral blood mononuclear cells accumulated 20-fold greater levels of GM-CSF mRNA and secreted comparably greater amounts of cytokine after transfection with hGM-AUGUA expression vectors versus hGM-AUUUA. hGM-AUGUA mRNA was fivefold more stable (t 1/2 = 95 minutes) than hGM-AUUUA mRNA (t 1/2 = 20 minutes), accounting for elevated steady-state levels. Transfection site extracts and serum samples obtained 24 hours after gene transfer of hGM-AUGUA cDNA into mouse skin contained greater than 32 ng/mL and 650 pg/mL of GM-CSF protein, respectively, compared with 0.33 ng/mL and less than 8 pg/mL for hGM-AUUUA cDNA. GM-CSF produced from mGM-AUGUA cDNA transfected into rat abdominal epidermis induced a profound neutrophil infiltrate. These data suggest a novel strategy for enhanced production of biologically active cytokines by normal cells after in vivo gene transfer.

Animals↗
PubMed1995

Assembly of a chromosomal replication machine: two DNA polymerases, a clamp loader, and sliding clamps in one holoenzyme particle. I. Organization of the clamp loader.

The gamma complex of DNA polymerase III holoenzyme, the replicase of Escherichia coli, couples ATP hydrolysis to the loading of beta sliding clamps onto primed DNA. The beta sliding clamp tethers the holoenzyme replicase to DNA for rapid and processive synthesis. In this report, the gamma complex has been constituted from its five different subunits. Size measurements and subunit stoichiometry studies show a composition of gamma 2 delta 1 delta' 1 1 chi 1 psi 1. Strong intersubunit contacts have been identified by gel filtration, and weaker contacts were identified by surface plasmon resonance measurements. An analogous tau complex has also been constituted and characterized; it is nearly as active as the gamma complex in clamp loading activity, but as shown in the fourth report of this series, it is at a disadvantage in binding the delta, delta', chi, and psi subunits when core is present (Xiao, H., Naktinis, V., and O'Donnell, M. (1995) J. Biol. Chem. 270, 13378-13383). The single copy subunits within the gamma complex provide the basis for the structural asymmetry inherent within DNA polymerase III holoenzyme.

Chromatography, High Pressure Liquid↗
PubMed1995

Assembly of a chromosomal replication machine: two DNA polymerases, a clamp loader, and sliding clamps in one holoenzyme particle. III. Interface between two polymerases and the clamp loader.

The nine-subunit DNA polymerase (Pol) III* coupled to its beta sliding clamp is a rapid and highly processive replicating machine. The multiple subunits are needed for the complicated task of duplicating the Escherichia coli chromosome. In this report, Pol III* was constituted from individual pure proteins, and its structure was studied. Constitution of the Pol III* particle requires an ordered addition of the subunits, and the final structure contains 14 polypeptides in the ratio alpha 2 epsilon 2 theta 2 tau 2 gamma 2 delta 1 delta' 1 chi 1 psi 1. The structure can be summarized as being composed of two core polymerases (alpha epsilon theta) held together by a dimer of tau and one gamma complex clamp loader (gamma 2 delta 1 delta' 1 chi 1 psi 1) for loading beta onto DNA. At the center of the structure, the related tau and gamma subunits form a heterotetramer upon which the two core polymerases and clamp loader proteins assemble. The single copy nature of the delta, delta', chi, and psi subunits confers a structural asymmetry with respect to the two polymerases, presumably for the different functions of replicating the leading and lagging strands.

Chromosomes, Bacterial↗
PubMed1995

In vivo cytokine gene transfer by gene gun reduces tumor growth in mice.

Implantation of tumor cells modified by in vitro cytokine gene transfer has been shown by many investigators to result in potent in vivo antitumor activities in mice. Here we describe an approach to tumor immunotherapy utilizing direct transfection of cytokine genes into tumor-bearing animals by particle-mediated gene transfer. In vivo transfection of the human interleukin 6 gene into the tumor site reduced methylcholanthrene-induced fibrosarcoma growth, and a combination of murine tumor necrosis factor alpha and interferon gamma genes inhibited growth of a renal carcinoma tumor model (Renca). In addition, treatment with murine interleukin 2 and interferon gamma genes prolonged the survival of Renca tumor-bearing mice and resulted in tumor eradication in 25% of the test animals. Transgene expression was demonstrated in treated tissues by ELISA and immunohistochemical analysis. Significant serum levels of interleukin 6 and interferon gamma were detected, demonstrating effective secretion of transgenic proteins from treated skin into the bloodstream. This in vivo cytokine gene therapy approach provides a system for evaluating the antitumor properties of various cytokines in different tumor models and has potential utility for human cancer gene therapy.

Animals↗
PubMed1995

Sequence, expression and transcriptional analysis of the coronafacate ligase-encoding gene required for coronatine biosynthesis by Pseudomonas syringae.

Pseudomonas syringae pv. glycinea PG4180 produces the chlorosis-inducing phytotoxin coronatine (COR), which consists of a polyketide component, coronafacic acid (CFA), ligated by an amide bond to coronamic acid (CMA), an ethylcyclopropyl amino-acid derived from isoleucine. We report the nucleotide sequence of a 2.37-kb region containing the coronafacate ligase-encoding gene (cfl) which is required for the amide linkage of CFA and CMA. The transcription start point for cfl was identified, and the Cfl protein was overproduced from the T7lac promoter in Escherichia coli. The deduced amino-acid sequence of Cfl showed homology to a variety of adenylate-forming enzymes which bind and hydrolyze ATP in order to activate their substrates for further ligation.

Amide Synthases↗
PubMed1995

Expression of functional GABAA receptors in neuroendocrine gastropancreatic cells.

Gastropancreatic neuroendocrine cells synthesize large amounts of gamma-aminobutyric acid (GABA). This amino acid neurotransmitter appears to be stored in and released from, vesicles similar to small synaptic vesicles. So far, the function of GABA in gastropancreatic, neuroendocrine cells has not been clarified. Previous work suggested that only pancreatic, glucagon-producing alpha 2 cells contain functional GABAA receptors. Using subunit-specific antibodies in sections of human antral mucosa, a human gastrinoma and rat pancreas, we show that expression of GABAA receptors is abundant in gastropancreatic, neuroendocrine cells. Using the patch-clamp technique in the whole-cell mode we demonstrate that both the rat insulinoma cell line RIN 38 and the amphicrine cell line AR42J express functional GABAA receptors, which are characterized by a relatively low benzodiazepine and Zn2+ sensitivity and by an insensitivity to the inverse benzodiazepine agonist 6,7-alpha-methoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM). In contrast to neurons, activation of GABAA receptors leads to a membrane depolarization. This depolarization presumably activates voltage-gated Ca2+ channels, resulting in an increase in cytosolic Ca2+ concentration, [Ca2+]i, as shown with the fluorimetric dye fura-2. The combination of GABA release, GABAA receptor activation and the [Ca2+]i increase could constitute an autocrine mechanism, modulating the release of hormones such as gastrin, insulin and somatostatin.

Animals↗
PubMed1995

Inhibition of beta A4 production by specific modulation of beta-secretase activity.

To study amyloid precursor protein (APP) processing we expressed different APP isoforms with and without the Swedish mutation and the membrane inserted C-terminal 100 residues of APP (SPA4CT) in the human neuroblastoma cell line SY5Y. We show that expression of the Swedish mutation results in a significant production of the amyloidogenic intermediate A4CT, which is further processed by gamma-secretase leading to an overproduction of beta A4. Treatment with methylamine and ammonium chloride, inhibitors interfering with intracellular transport mechanisms, inhibits beta-secretase activity without influencing the physiological APP cleavage by alpha-secretase activity. By expressing SPA4CT, we demonstrate that secretion, but not generation, of beta A4 from SPA4CT is inhibited by methylamine resulting in intracellular beta A4. This provides experimental evidence for the intracellular localization of gamma-secretase activity and beta A4 generation.

Ammonium Chloride↗
PubMed1995

Transfection of a murine fibrosarcoma with intercellular adhesion molecule-1 enhances the response to adoptive immunotherapy.

BACKGROUND: Increasing the ability of antitumor effector cells to leave the vasculature and gain access to tumor cells may improve therapeutic efficacy. We undertook this study to determine whether increased expression of intercellular adhesion of molecular-1 (ICAM-1) by gene transfection would result in an improved response to adoptive immunotherapy in vivo. METHODS: C57BL/6 mice received 1 x 10(6) tumor cells on day 0. Tumor cells examined were MCA-105 (parental), NeoR (MCA-105 transfected with the neomycin resistance gene), or Clones 81 or 149 (MCA-105 cotransfected with NeoR and the gene for ICAM-1 and highly express ICAM-1). Animals were treated by use of no treatment, interleukin-2 alone (days 10 through 14), hyperthermia alone (days 10 and 13), or interleukin-2 + hyperthermia, and tumor growth was reported as a ratio to size on day 10. In vitro cytotoxicity was assayed by using murine lymphokine-activated killer cells. RESULTS: Tumors transfected with ICAM-1 and treated with hyperthermia + immunotherapy grew significantly (p < 0.05) slower (mean, 0.78 +/- 0.16 on day 19) than parental tumor (size, 1.35 +/- 0.22) or tumor cells transfected with NeoR alone (1.21 +/- 0.19). Tumors containing both MCA-105 and Clone 81 treated with hyperthermia + immunotherapy grew significantly slower (1.58 +/- 0.49 on day 19, p < 0.05) than untreated Clone 81 (2.38 +/- 0.46) or treated MCA-105 (2.49 +- 0.29) but more rapidly than treated Clone 81 (1.18 +/- 0.08), suggesting a paracrine efect for ICAM-1. CONCLUSIONS: These findings show that increased expression of ICAM-1 by tumor cells results in a significant increase in antitumor efficacy of combined interleukin-2 and hyperthermia in a murine model. Although the mechanism has yet to be elucidated, modulation of cellular adhesion may play a role in the therapeutic efficacy of cellular immunotherapy.

Animals↗
PubMed1995

Relationship between psychiatric disease and neuropsychological impairment in HIV seropositive individuals.

Neuropsychological impairment and DSM-III-R Axis I psychiatric diagnoses were evaluated in a heterogenous group of HIV seropositive individuals and seronegative individuals with similar risk factors for HIV infection. Neuropsychological and psychiatric disorders were common in the HIV seropositive group, but there were no relationships between these two aspects of neuropsychiatric dysfunction in seropositive patients. Results indicate that psychiatric disorders in HIV seropositive individuals tend to predate infection and decrease over time following knowledge of seroconversion, suggesting that they are primarily a function of psychosocial factors. Neuropsychological disorders are specific to HIV seropositive patients and tend to increase over time following seroconversion, suggesting that they are due to neurological effects of HIV-infection.

AIDS Dementia Complex↗
PubMed1995

Detection of surfaces in structure from motion.

In 3 experiments the authors examined the effects of surface shape on the detection of surfaces in structure from motion displays. In experiment 1 they found the number of points necessary to distinguish simple quadratic surfaces from points randomly placed in a volume to be low, in the range of 4-6 points. In experiment 2 they found that a hyperbolic shape is harder to detect than an arch or parabolic shape and that more curved shapes are easier to detect than less curved shapes. In experiment 3 they found that detection of sinusoidal shapes increases with increasing amplitude and decreasing frequency, confirming previous results with motion parallax displays. This pattern of results is consistent with a model in which triangular surface patches are fit through adjacent triples of points and displays with smaller mean orientation change between neighboring triangles are judged to be surface displays.

Humans↗
PubMed1995

The AXR1 and AUX1 genes of Arabidopsis function in separate auxin-response pathways.

The recessive mutations aux1 and axr1 of Arabidopsis confer resistance to the plant hormone auxin. The axr1 mutants display a variety of morphological defects. In contrast, the only morphological defect observed in aux1 mutants is a loss of root gravitropism. To learn more about the function of these genes in auxin response, the expression of the auxin-regulated gene SAUR-AC1 in mutant and wild-type plants has been examined. It has been found that axr1 plants display a pronounced deficiency in auxin-induced accumulation of SAUR-AC1 mRNA in seedlings as well as rosette leaves and mature roots. In contrast, the aux1 mutation has a modest effect on auxin induction of SAUR-AC1. To determine if the AUX1 and AXR1 genes interact to facilitate auxin response, plants which are homozygous for both aux1 and axr1 mutations have been constructed and characterized. The two mutations are additive in their effects on auxin response, suggesting that each mutation confers resistance by a different mechanism. However, the morphology of double mutant plants indicates that there is an inter-action between the AXR1 and AUX1 genes. In mature plants, the aux1-7 mutation acts to partially suppress the morphological defects conferred by the axr1-12 mutation. This suppression is not accompanied by an increase in auxin response, as measured by SAUR-AC1 expression, suggesting that the interaction between the AUX1 and AXR1 genes is indirect.

2,4-Dichlorophenoxyacetic Acid↗