Search PubMed⌕ Search

Biomedical subjects

F M Davis

Publications and source records attributed to F M Davis.

At least 37 records · Page 2Linked to original sources

A new isoform of human membrane-bound IgE.

The epsilon-chain of membrane-bound IgE on the surface of B lymphocytes is known to contain a membrane-anchoring peptide segment that is encoded by two membrane exons, me.1 and me.2. In analyzing pertinent segments in mRNA from human IgE-expressing B cells by using PCR methods and Northern blotting analyses, we have identified three species of mRNA of epsilon-chain with variations in the splicing of the membrane exons. The conventional species (m/s) contains the predicted me.1 and me.2; species m/1 harbors 156 extra nucleotides 5' of me.1 with unaltered reading frame; species s/t lacks me.1 and hence the segment encoding the hydrophobic transmembrane stretch and contains a shifted me.2 reading frame. Rabbit antibodies, which were prepared by immunization using a peptide of 36 amino acid residues representing an encoded segment unique to mRNA species m/l, could specifically bind to human IgE-expressing B cell lines and react with an epsilon-chain on Western immunoblots. These results indicate that there exists a previously unidentified isoform of human membrane-bound IgE.

Amino Acid Sequence↗

Lidocaine for the prevention of pain due to injection of propofol.

Propofol has a high incidence of pain with injection, particularly into small veins. We sought to determine whether concomitant administration of lidocaine could prevent this pain. In a randomized double-blind trial, 368 women were allocated to one of four groups to receive 19 mL of propofol mixed with either 1 mL of 0.9% saline, 1 mL of 0.5% (5 mg) lidocaine, 1 mL of 1% (10 mg) lidocaine, or 1 mL of 2% (20 mg) lidocaine. The pain of injection was scored as none, mild, moderate, or severe. There was a significant reduction in the overall incidence of pain from 73% with saline to 32% with 20 mg lidocaine. A highly significant negative dose-response relationship between the dose of lidocaine and the severity of pain was demonstrable, both at induction of anesthesia and as recalled in the recovery room (P less than 0.001 for both). Lidocaine (20 mg IV) will significantly reduce the incidence and severity of pain with propofol injection, but about 6% of patients will still suffer unpleasant pain if the dorsum of the hand is used.

Adult↗

MPM-12: a monoclonal antibody that predominantly stains mitotic cells and recognizes a protein kinase.

The monoclonal antibody MPM-12, raised by using partially purified extract of mitotic HeLa cells as the immunogen, preferentially stains the cytoplasm of mitotic cells by indirect immunofluorescence without exhibiting any species specificity. On immunoblots, MPM-12 recognizes three bands, of 155, 88, and 68 kDa, in mitotic HeLa cell extract but only the 68-kDa band in interphase cell extract. The 68-kDa band seems to be associated with chromatin while the other two are not. All three MPM-12 reactive peptides are phosphorylated, and the phosphorylation seems to be required for MPM-12 reactivity. The MPM-12 immunocomplexes exhibit autophosphorylating and histone H1 kinase activity.

Antibodies, Monoclonal↗

Delayed hyperbaric oxygen therapy for carbon monoxide intoxication--two case reports.

The severity of signs and symptoms following carbon monoxide intoxication often does not relate to admission carboxyhaemoglobin levels. Two cases are presented with severe neurological impairment despite carboxyhaemoglobin levels of 2% and 1.7% on admission to hospital, who responded well to hyperbaric oxygen therapy. In one case, symptoms recurred several days later, but responded to further hyperbaric oxygen therapy. The role of and possible mechanism of action of late hyperbaric oxygen treatment in carbon monoxide intoxication are discussed.

Ataxia↗

Transfectomas expressing both secreted and membrane-bound forms of chimeric IgE with anti-viral specificity.

Because of the lack of a cell line expressing on surface and secreting human IgE of known Ag specificity, the construction of a transfectoma line possessing such properties would be useful for studying the roles of surface IgE and the effects of anti-IgE antibodies on IgE-producing B cells. Toward this goal, the human genomic DNA segment encompassing the two exons encoding the membrane anchor peptide of epsilon-chain and their flanking regions was sequenced. Hybrid epsilon and kappa genomic DNA comprising the C regions of human epsilon- and kappa-chains and the H and L chain V regions of the murine mAb BAT123, which reacts with the gp120 envelope protein of HIV-1, were constructed. Mammalian expression vectors containing these fusion genes were used to transfect murine myeloma Sp2/0 cells, and transfectants stably expressing on surface and secreting into culture medium chimeric IgE were obtained. The chimeric IgE showed identical Ag-binding properties as the murine mAb BAT123. Acting in concert with the specific peptide Ag polyvalently coupled to a protein carrier, the chimeric antibody could induce histamine release from human blood basophils. These results demonstrate the potential utility of the transfectoma cells and the chimeric IgE in studying the roles of membrane-bound IgE and effects of anti-IgE antibodies on IgE-producing B cells.

Amino Acid Sequence↗

An epitope on membrane-bound but not secreted IgE: implications in isotype-specific regulation.

Immunoglobulins (Igs) on the surface of B lymphocytes are isotype-specific immunological markers of the B-cell subsets expressing them. Since these membrane-bound Igs (mIgs) are antigen receptors, their interaction with antibodies could be explored for modulating the activity of specific B-cell subsets. Targeting mIgs by antibodies in vivo, however, has not been feasible because of the presence of Igs in the circulation and the frequent association of Igs with various cell types via Fc receptors. To circumvent these problems, we proposed that the extracellular portions of the membrane-anchoring segments of the heavy chains of mIgs, referred to as "mIg isotype-specific" or "migis" peptides, may provide the antigenic sites for the isotype-specific targeting of B cells in vivo. Here we describe the exemplary development of monoclonal antibodies (mAbs) recognizing this unique epitope of mIgE.

Amino Acid Sequence↗

Monoclonal antibodies specific for human IgE-producing B cells: a potential therapeutic for IgE-mediated allergic diseases.

Monoclonal antibodies (MAbs) specific for surface antigens of lymphocytes are being used to target and deplete tumorous or normal lymphocytes in vivo. Here, we report evidence for the existence of antigenic epitopes on IgE that are accessible on IgE-secreting B cells but not on other cells bearing IgE. Among 42 murine MAbs specific for human IgE, two were shown by fluorescence flow cytometric analyses to bind to IgE-secreting cell lines but not to IgE bound to high-affinity IgE.Fc receptors (Fc epsilon RI) on basophils or low-affinity IgE receptors (Fc epsilon RII) on other cell types. Neither could they induce histamine release from basophils of various donors even under very permissive conditions. These antibodies may be useful for targeting IgE-secreting B cells in patients suffering from IgE-mediated allergies.

Animals↗

Distribution of phosphorylated spindle-associated proteins in the diatom Stephanopyxis turris.

Mitotic spindles isolated from the diatom Stephanopyxis turris become thiophosphorylated in the presence of ATP gamma S at specific locations within the mitotic apparatus, resulting in a stimulation of ATP-dependent spindle elongation in vitro. Here, using indirect immunofluorescence, we compare the staining pattern of an antibody against thiophosphorylated proteins to that of MPM-2, an antibody against mitosis-specific phosphoproteins, in isolated spindles. Both antibodies label spindle poles, kinetochores, and the midzone. Neither antibody exhibits reduced labeling in salt-extracted spindles, although prior salt extraction inhibits thiophosphorylation in ATP gamma S. Furthermore, both antibodies recognize a 205 kd band on immunoblots of spindle extracts. Microtubule-organizing centers and mitotic spindles label brightly with the MPM-2 antibody in intact cells. These results show that functional mitotic spindles isolated from S. turris are phosphorylated both in vivo and in vitro. We discuss the possible role of phosphorylated cytoskeletal proteins in the control of mitotic spindle function.

Antibodies↗

Leg blood flow during total hip replacement under spinal or general anaesthesia.

Calf blood flow was studied using venous occlusion impedance plethysmography during 122 total hip arthroplasties. Patients were randomly allocated to receive spinal or general anaesthesia. Blood flow was measured nine times perioperatively. In the non-surgical leg, mean blood flow rose by over 50% in both groups following anaesthetic induction, remaining significantly elevated with spinal but falling back gradually to baseline with general anaesthesia. In the surgical leg, surgical manipulations produced marked falls in flow in many patients, particularly with femoral component insertion. If this occurred, hyperaemia was commonly seen with spinal anaesthesia but rarely with general anaesthesia once the joint was relocated. Venous outflow resistance rose slightly during anaesthesia in both groups, more so with general anaesthesia. In the surgical leg, marked rises occurred with surgical manipulations, but resistance fell abruptly once the joint was relocated. No clear relationship between these observations and the occurrence of deep vein thrombosis postoperatively was established.

Anesthesia, General↗

Preoperative transdermal hyoscine for the prevention of postoperative nausea and vomiting.

In a randomised, double-blind study, a transdermal patch containing either hyoscine or placebo was applied post-auricularly in 190 adult patients under 65 year old, seven to twelve hours prior to their undergoing minor orthopaedic or plastic surgery under thiopentone/nitrous oxide/halothane general anaesthesia. In the first 24 hours after surgery 34% of patients vomited. The incidence of nausea (31% vs 54%; P less than 0.01) and the number of episodes of vomiting (66 vs 125; P less than 0.05) during the first 24 hours were significantly less with hyoscine than with placebo. The hyoscine group required fewer doses of antiemetic than the placebo group (12 vs 27; P less than 0.05). Side-effects were mild, the only difference between the two groups being the frequency of dry mouth immediately preoperatively. No differences were seen in the second 24 hours after surgery. We conclude that transdermal hyoscine is moderately effective in reducing the occurrence of postoperative nausea and vomiting following minor surgery.

Administration, Cutaneous↗

Mitosis-specific monoclonal antibodies block cleavage in amphibian embryos.

By microinjecting monoclonal antibodies that bind specifically to mitotic and meiotic cells of a variety of species, we studied the biological activity of antigens recognized by these antibodies. The antibodies recognize a family of phosphoprotein antigens that are found throughout the cytoplasm of mitotic cells and particularly at microtubule organizing centers, including centrosomes and kinetochores. Their binding is dependent on phosphorylation of the polypeptides. Immunoglobulins were introduced into Xenopus laevis and Rana pipiens oocytes or cleaving embryos using glass micropipettes. The ability of the antibody-injected oocytes to undergo mitosis or meiosis was compared with those injected with control mouse immunoglobulins. The antibodies failed to block chromosome condensation and germinal vesicle breakdown in progesterone-treated oocytes. However, functional mitotic spindles were not assembled in cleavage stage frog embryos injected with antibodies. In vitro, the binding of the antibodies to the antigens inhibited the dephosphorylation of the antigens by alkaline phosphatase. The antibody binding to the activated microtubule organizing centers (MTOC) seems to block not only the nucleation of microtubules and the organization of the mitotic spindle, but also the dephosphorylation of proteins associated with the MTOC that normally occurs at the mitosis-G1 transition.

Animals↗

Deep vein thrombosis after total hip replacement. A comparison between spinal and general anaesthesia.

The effect of hypobaric spinal anaesthesia or narcotic-halothane-relaxant general anaesthesia on the incidence of postoperative deep vein thrombosis was studied in 140 elective total hip replacements in a prospective randomised manner. Deep vein thrombosis was diagnosed using impedance plethysmography and the 125I fibrinogen uptake test, combined, in selected cases, with ascending contrast venography. The overall incidence of deep vein thrombosis was 20%. Nine patients (13%) developed deep vein thrombosis in the spinal group and nineteen (27%) in the general anaesthetic group (p less than 0.05). The incidences of proximal thrombosis and of bilateral thrombi were also less with spinal anaesthesia than with general anaesthesia. It is concluded that spinal anaesthesia reduces the risks of postoperative thromboembolism in hip replacement surgery. The presence of varicose veins, being a non-smoker and having a low body mass index were associated with an increased incidence of deep vein thrombosis.

Aged↗

Movement and segregation of kinetochores experimentally detached from mammalian chromosomes.

The kinetochore is a specialized structure at the centromere of eukaryotic chromosomes that attaches chromosomes to the mitotic spindle. Recently, several lines of evidence have suggested that kinetochores may have more than a passive role in the movement of chromosomes during mitosis and meiosis. Kinetochores seem to attract and 'capture' microtubules that grow from the spindle poles and microtubules may lengthen or shorten by the addition or subtraction of tubulin subunits at their kinetochore-associated ends. An attractive hypothesis is that kinetochores function as 'self-contained engines running on a microtubule track'. Here, we show that kinetochores can be experimentally detached from chromosomes when caffeine is applied to Chinese hamster ovary cells that are arrested in the G1/S phase of the cell cycle. The detached kinetochore fragments can still interact with spindle microtubules and complete all the mitotic movements in the absence of other chromosomal components. As these cells enter mitosis before DNA synthesis is completed, chromosome replication need not be a prerequisite for the pairing, alignment and segregation of kinetochores.

Animals↗

Selective inhibition of human ribosomal gene transcription by the morpholinyl anthracyclines cyanomorpholinyl- and morpholinyldoxorubicin.

Upon incubation of cultured mammalian cells with the new anthracycline analogues cyanomorpholinyldoxorubicin and morpholinyldoxorubicin, nucleoli irreversibly segregate into their substructures which form individual portions of the nucleolar mass and characteristic electron-dense components adjacent to the nucleolonema; these changes in nucleolar ultrastructure are similar to those produced by actinomycin D (AMD). In the present study we have examined the effects of anthracycline analogues on RNA synthesis, localization of RNA polymerase I in situ, and activity of RNA polymerases in vitro, and compared these effects with those of the parent compound doxorubicin (DOX) and AMD. The results show that, following treatment with cyanomorpholinyldoxorubicin, morpholinyldoxorubicin, and AMD, but not DOX, RNA polymerase I-containing transcription complexes were reduced, reflecting the transcriptional activity of the rRNA genes. The residual RNA polymerase-containing entities were redistributed into cap-like aggregates at the nucleolar periphery. Within 30 min of exposure to cyanomorpholinyldoxorubicin, morpholinyldoxorubicin, and AMD, but not DOX, a 75-90% inhibition of RNA polymerase I activity in situ and in vitro was observed. At this early time there was no significant inhibition of nucleoplasmic RNA labeling in situ or RNA polymerases II and III activities in vitro. At later times following reincubation in drug-free medium, inhibition of all three polymerases was observed. Impairment of RNA synthesis appeared to result from drug interaction with the DNA template rather than an interaction with RNA polymerase I itself. We conclude that the morpholinyl derivatives of DOX are preferential inhibitors of ribosomal gene transcription and that they may have a mechanism of action similar to that of AMD on rRNA synthesis.

Dactinomycin↗