Biomedical subjects
Publications and source records attributed to J H Phillips.
A new method for the injection of anesthetics is discussed for the tumescent technique of local anesthesia for liposuction. A 12-gauge 30-cm needle with numerous injection ports is used with an injection gun. The technique requires minimal effort and has been found to be a more rapid method of injecting large volumes of fluid than previous systems.
Pertussis toxin stimulates both basal and nicotine-evoked catecholamine secretion from intact bovine adrenal chromaffin cells, as well as Ca2(+)-evoked release from permeabilized cells. Tetanus toxin inhibits all these effects; it reduces the secretion of intact cells treated with pertussis toxin to the basal level, and decreases by about 50% Ca2(+)-evoked release from permeabilized cells whether or not previously stimulated by pertussis toxin.
The majority of human NK cells express low affinity IgG Fc receptors (CD16+), whereas a minor subset of NK cells lack Fc receptor expression (CD16-). In contrast to CD16+ NK cells that express only p75 IL-2 receptors, CD16- NK cells constitutively co-express both p75 and p55 IL-2 receptors in vivo and preferentially respond to low concentrations of IL-2 with increased cytolytic activation and proliferation. Scatchard analysis demonstrated the presence of approximately 1,200 high affinity (approximately 25 pM kD) and approximately 9,600 intermediate affinity (approximately 2 nM kD) IL-2 receptors on CD16- NK cells. CD16+ NK cells expressed only a single intermediate affinity IL-2 receptor of approximately 1.9 nM kD (approximately 9,000 sites per cell). The IL-2 binding data thus substantiated the phenotypic and functional studies and definitively show that the differential responsiveness of CD16- and CD16+ NK cells to IL-2 is manifested through different affinity IL-2 receptors.
Collapse of the zygomatic arch following trauma results in inadequate anteroposterior projection of the zygomatic body and an increase in facial width. Accurate assessment of the position of the zygomatic arch in relation to the cranial base posteriorly and the midface anteriorly is the key to the acute repair of complex midfacial fractures and the secondary reconstruction of posttraumatic deformities of the orbitozygomaticomaxillary complex. Loss of projection of the zygomatic arch may occur with injuries confined to the orbitozygomaticomaxillary region or in association with complex midfacial fractures. A safe anatomic approach to the zygomatic arch allows exact anatomic restoration of the zygomatic arch using miniplates and screws and results in the reconstruction of an outer facial frame with a correct anteroposterior projection and facial width. The zygomatic arch injury is diagnosed using axial CT scanning. Three-hundred and seventeen arches have been exposed through a coronal incision following acute trauma and 47 arches have been exposed in patients requiring late correction of a posttraumatic orbitozygomaticomaxillary deformity. Permanent palsy to the frontal branch of the facial nerve has occurred in one patient following the exact definition of the anatomy of this region.
This paper investigates the relationship between bone resorption, the process of bone revascularization, and graft fixation. Vital staining techniques and microangiography were used to study the extent of graft revascularization of fixed and nonfixed endochondral (rib) and membranous (skull) onlay bone grafts in 20 adult sheep mandibles bilaterally. This assessment was carried out at 2 and 20 weeks postoperatively. Sequential fluorochrome staining was performed to examine the pattern of new bone deposition. Fixation was achieved using the lagscrew technique. At 2 weeks, membranous bone demonstrated a greater area of graft revascularization if fixed than if the graft was not fixed. The opposite result was seen for endochondral grafts, where nonfixed grafts showed a greater area of revascularization than fixed grafts. At 20 weeks, all bone that was present was fully vascularized. The inconsistencies in the results on the relationship between fixation and revascularization for membranous and endochondral grafts in the early stages of healing (2 weeks) suggest that although revascularization is a necessary precondition for bone resorption and deposition, biomechanical and structural factors may be a more satisfactory explanation for the differences observed in the maintenance of bony volume.
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CD16 is a low-affinity immunoglobulin G (IgG) Fc receptor that is expressed on natural killer (NK) cells, granulocytes, activated macrophages, and some T lymphocytes. Two similar genes, CD16-I and CD16-II, encode membrane glycoproteins that are anchored by phosphatidylinositol (PI)-glycan and transmembrane polypeptides, respectively. The primary structural requirements for PI-linkage were examined by constructing a series of hybrid cDNA molecules. Although both cDNA's have an identical COOH-terminal hydrophobic segment, CD16-I has Ser203 whereas CD16-II has Phe203. Conversion of Phe to Ser in CD16-II permits expression of a PI-glycan-anchored glycoprotein, whereas conversion of Ser to Phe in CD16-I prevents PI-glycan linkage.
Natural killer (NK) cells are a subset of lymphocytes that mediate major histocompatibility complex (MHC)-nonrestricted cytotoxicity against tumours and virus-infected cells and secrete numerous cytokines on activation. NK cells are distinct from mature T lymphocytes, because they do not rearrange or productively transcribe T-cell receptor alpha-, beta-, gamma- or delta-chain genes and do not express the CD3 gamma- or delta-subunits. But recent studies indicate that NK cells do express CD3 zeta, co-associated with other membrane proteins. Here we report that CD16, the receptor for the Fc (constant) region of IgG, specifically associates with the CD3 zeta homodimer on the membrane of human NK cells, and that co-transfection with CD3 zeta complementary DNA permits expression of a transmembrane-linked CD16 complex on COS-7 cells. These findings indicate that CD3 zeta can co-associate with membrane receptors of diverse cell types and function as a common structure for signal transduction.
In the present study we have identified and characterized three subpopulations of peripheral blood NK cells based on the surface expression of CD56 and CD16. We have designated these subsets CD16neg, CD16dim, and CD16bright according to the relative surface density of CD16. The CD16bright subset comprised about 10% to 15% of PBL, whereas the CD16dim and CD16neg subsets comprise less than 1% of the total lymphocytes. A detailed characterization of these subsets revealed both similarities and differences. The three subsets shared a great deal of phenotypic similarity, expressing CD2, CD7, CD11b, CD38, CD45R, CD18, and the p75 IL-2R on the majority of the cells in each subset. There were, however, several prominent phenotypic differences, particularly in the expression of CD57, CD11c, CD44, CD25, Leu-8, L263, and L265. The CD16neg cells were morphologically large agranular lymphocytes and demonstrated low levels of non-MHC restricted cytolysis of NK-sensitive tumor lines. The CD16dim and CD16bright subsets were large granular lymphocytes and revealed potent cytotoxicity against NK-sensitive targets. All subsets demonstrated IL-2-dependent activation and proliferation; however, the CD16dim and CD16neg subsets were preferentially responsive to very low concentrations of rIL-2. Although rIL-4 effectively inhibited the IL-2-induced cytolytic activation of all three NK cell subsets, only the CD16bright cells showed rIL-4 inhibition of IL-2 dependent proliferation. Cytokine transcription was also differentially regulated in the NK cell subsets after rIL-2 activation. Although TNF-alpha was equally transcribed in each subsets, IFN-gamma and serine protease-HF were preferentially transcribed in the CD16bright NK cells. Based on these results, we propose that these NK cell subsets represent portions of the NK cell differentiation pathway present in the peripheral blood.
The CD69 (Leu-23) activation Ag is a phosphorylated 28 to 32-kDa disulfide-linked homodimer that is rapidly induced after lymphocyte activation. CD69 is not present on the surface of peripheral blood resting T cells, but is constitutively expressed by CD3bright thymocytes. Activation of protein kinase C (PKC) by stimulation of the TCR/CD3 or by phorbol esters directly induces CD69 expression on T cells. In the attempt to elucidate the function of CD69 we investigated the ability of the CD69 glycoprotein to transmit an activation signal. Cross-linking of CD69 by mAb induced a prolonged elevation of intracellular [Ca2+], mostly due to an influx of extracellular Ca2+. This signal alone was unable to effectively activate PKC. When PKC was simultaneously activated by PMA, stimulation of CD69 induced IL-2 and IFN-gamma gene expression, enhancement of CD25 expression, and ultimately IL-2-dependent T cell proliferation. Both CD4+ and CD8+ peripheral T cells responded to CD69-mediated activation. Stimulation of CD69 induced proliferation of thymocytes as well as peripheral T cells, but both required independent PKC activation by PMA. Cyclosporin A, which does not prevent PKC-induced CD69 expression, completely suppressed CD69-induced IL-2 and IFN-gamma gene expression. Although the signal delivered by the CD69 initiates T cell proliferation, it is unable to trigger cytotoxicity programs in CD69+-activated T cells or T cell clones.
The IL-2R is composed of at least two subunits, the p55 (CD25/Tac) and p75 glycoproteins. The p75 IL-2R is expressed preferentially on resting human peripheral blood NK cells, but is not detected on substantial proportions of T and B lymphocytes, monocytes, or granulocytes. Anti-p75 IL-2R mAb substantially inhibits the early events associated with NK cell activation by IL-2, including inhibition of cytotoxic activity and induction of the CD69 early activation antigen. While anti-p55 IL-2R mAb alone failed to substantially inhibit the initial events of IL-2 stimulation, maximal inhibition of IL-2-induced cytotoxicity and proliferation was achieved by combining both anti-p55 IL-2R and anti-p75 IL-2R. Collectively, results from the present studies directly implicate the p75 IL-2R as the structure predominantly responsible for IL-2 activation of NK cells.
Neural cell adhesion molecule (N-CAM) is a membrane glycoprotein expressed on neural and muscle tissues that is involved in homotypic adhesive interactions. We have demonstrated that N-CAM also is expressed on hematopoietic cells, and is recognized by the anti-Leu-19 mAb. Leu-19 is preferentially expressed on NK cells and T lymphocytes that mediate MHC-unrestricted cytotoxicity, but is also present on some myeloid leukemia cell lines. On NK cells, T cells, the KG1a.5 hematopoietic cell line, and a neuroblastoma cell line, Leu-19 is a approximately 140-kD polypeptide with N-linked carbohydrates and abundant sialic acid residues. Sequential immunoprecipitation and peptide mapping demonstrated that the Leu-19 and N-CAM molecules expressed on leukocyte and neuroblastoma cell lines are similar structures. These findings suggest that the Leu-19 antigen on leukocytes may be involved in cell adhesion, analogous to the function on N-CAM on neural cells.
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We have measured the contents of Na+ and K+ in isolated chromaffin granules. Total contents varied between 227 and 283 nmol/mg of protein, equivalent to matrix concentrations of 53-66 mM. The value found depended on the isolation buffer used, and the ratio of the two ions reflected the composition of the buffer. We then measured the free concentration of each of these ions, and of Ca2+, in the matrix, by using a null-point method with acridine-fluorescence quenching. This monitored H+ fluxes induced by an ionophore in the presence of known concentrations of the ion in the supporting medium. In contrast with organic constituents of the matrix, which have low activity coefficients, Na+ and K+ were found to have activity coefficients around 0.8 Ca2+, however, was strongly bound: its free concentration was only 0.03% of the total.
The Leu 23 Ag is a phosphorylated 28 to 32-kDa disulfide-linked homodimer expressed on the surface of activated T cells, B cells, and NK cells. Resting, unstimulated peripheral blood T cells are Leu 23-, but 20 to 30% of normal thymocytes constitutively express Leu 23. Triggering of the CD3/TCR complex by mAb induced the expression of Leu 23 within 2 h after stimulation. Leu 23 was still detectable on the cell surface 48 h after the stimulus was removed. The induction of Leu 23 strictly correlated with the extent of CD3/TCR cross-linking on the surface of T cells. Anti-CD3-coupled beads, but not soluble IgG or IgM mAb, were able to induce Leu 23 expression on peripheral blood T cells. Soluble anti-CD3 mAb were sufficient to induce Leu 23 on the Jurkat T cell line, but a direct relationship was found between CD3 cross-linking valency and Leu 23 expression. RNA and protein synthesis were both required for Leu 23 induction. Moreover, only CD3/TCR-mediated signals able to stimulate PKC and maintain elevated intracellular calcium ion concentration for greater than 60 min resulted in Leu 23 induction. These findings suggest that the extent of CD3/TCR cross-linking influences the persistence of elevated intracellular calcium ion concentration and PKC activation, and that this in turn determines the commitment of the T cell to activate gene transcription programs necessary for Leu 23 expression. Induction of Leu 23 represented the first detectable cell surface marker after triggering via the CD3/TCR and thus provides an ideal indicator to monitor the very early events in T cell activation.
Chromaffin granules, the secretory vesicles of the adrenal medulla, have a Na+/H+ exchange activity in their membranes which brings their proton gradient into equilibrium with a Na+ gradient. This explains why Na+ is mildly inhibitory to amine transport (which is driven by the H+ gradient) The activity can be demonstrated by using accumulation of 22Na+ in response to a pH gradient that is either imposed by diluting membrane 'ghosts' into alkaline media, or generated by ATP hydrolysis. It can also be monitored indirectly by fluorescence measurements in which the pH inside 'ghost' is monitored by quenching of a fluorescent weak base. This method has been used to monitor Na+ entry into acid-loaded 'ghosts' of H+ entry into methylamine accumulation. The exchanger appears to be reversible and non-electrogenic, with a stoichiometry of 1:1. Using an indirect assay we measured an apparent Km for Na+ of 4.7 mM, and a Ki for amiloride, a competitive inhibitor, of 0.26 mM. Direct assays using 22Na+ suggested a higher Km. Ethylisopropylamiloride was not inhibitory.
CD16 is an IgG Fc receptor that is predominantly expressed on human natural killer (NK) cells and granulocytes. The CD16 antigen expressed on neutrophils is a 50 to 70-kDa glycoprotein attached to the plasma membrane by a phosphatidylinositol-glycan linkage that is susceptible to cleavage by phosphatidylinositol-specific phospholipase C (PIPLC). By contrast, treatment of NK cells with PIPLC did not cleave CD16 from the cell surface. Biochemical analysis of the deglycosylated CD16 revealed a substantial difference in the core polypeptides expressed by NK cells and granulocytes. Culture of radiolabeled NK cells resulted in the spontaneous release of a CD16 molecule, smaller than the membrane form of CD16. These findings demonstrate that structural differences exist in the CD16 antigens expressed on NK cells and granulocytes that may potentially influence their functional activities.
The neurosurgeon and plastic surgeon are increasingly called upon to manage the care of patients with combined injuries of the cranium and face. The authors briefly review the pathogenesis and classification of craniofacial fractures and outline historical approaches to them. Current principles of management are then discussed. Experience with 167 patients is presented with emphasis on surgical technique, the sequence of repair and early primary reconstruction. The controversial issue of fontal sinus fracture repair is addressed. The authors favour preservation of the frontal sinus cavity, where possible, and do not obliterate the nasofrontal duct. With injuries to the floor of the sinus, the base of the sinus and frontonasal duct are sealed with bone graft and a vascularised soft tissue flap and the sinus is cranialised. Immediate bone grafts, using split skull or rib, are used to reconstruct areas of bony destruction or loss. Ninety-eight patients required 402 grafts. Immediate bone grafting resulted in few complications and low incidence of secondary deformities needing correction.