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Biomedical subjects

J M Robinson

Publications and source records attributed to J M Robinson.

At least 109 records · Page 6Linked to original sources

Detection of diaminobenzidine reactions using scanning laser confocal reflectance microscopy.

We used scanning laser confocal microscopy to visualize sites of peroxidatic activity as detected by the diaminobenzidine (DAB) reaction. Imaging was achieved by employing the reflectance mode of this instrument. Intense reflectance was detected after DAB localization of endogenous granule-associated myeloperoxidase in neutrophils and of the exogenous tracer horseradish peroxidase in mouse oocytes. Detection of DAB reaction products with confocal reflectance microscopy will probably be an important addition to the utility of this cytochemical technique.

3,3'-Diaminobenzidine↗

Morphology of the maxillary lateral incisor in cases of unilateral impaction of the maxillary canine.

Two groups of subjects who demonstrated unilateral maxillary canine impaction were examined, a Caucasian group and an Oriental group. It was found that there are significant differences between the groups for sex of patients seeking treatment, site of impaction (buccal/palatal), and arch form. The Oriental group was significantly more crowded in the canine area than the Caucasian group. In almost all cases the dimensions of the lateral incisor on the side of impaction were smaller than the non-impacted side, although five subjects from the Caucasian female sub-group did not conform to this pattern, and for the Oriental male sub-group the mesiodistal crown width was larger on the impacted side.

Adolescent↗

Modulation of murine hematopoiesis in vivo with recombinant murine interleukin-1.

Interleukin-1 (IL-1), a cytokine, primarily produced by monocytes, is the molecule involved in mediating many of the body's responses associated with infection and inflammation. More recently, IL-1 has been shown to sustain elevated levels of circulating granulocytes, stimulate the production of granulocyte-macrophage colony stimulating factors (CSFs) in vitro, increase plasma levels of CSF, and act synergistically with CSFs to increase the number of granulocyte-macrophage progenitors (colony-forming units) (CFU-GM) in vitro. The purpose of this study was to investigate the effect of murine IL-1 on steady-state hematopoiesis in vivo. C3H/HeJ or its normal littermate C3H/HeN male mice were administered either murine recombinant IL-1 at 45, 50, 200, 225, or 900 units (0.0125-0.25 micrograms)/animal, or 200 units (0.05 micrograms) of semipurified IL-1 derived from P388D1 cell culture supernatants. Because one of the responses to IL-1 is increased prostaglandin (PG) production and with the known activity of PGs on hematopoiesis, additional studies incorporated the cyclooxygenase inhibitor indomethacin (IM) (10 mg/kg body weight). In order to study the effect of IL-1 in vivo on pluripotential progenitors (CFU-S), IL-1 was compared with recombinant murine GM-CSF (50, 200, and 900 units; 0.0125, 0.05, and 0.25 micrograms). Control groups consisted of animals receiving either lipopolysaccharide (0.5 mg/kg body weight) or phosphate-buffered saline where appropriate. After 24 h, animals were sacrificed, and their peripheral blood indices and stem cell content of both bone marrow and spleen were evaluated for various committed hematopoietic progenitors: CFU-GM, CFU-Meg, CFU-E, BFU-E, and CFU-DG. Circulating neutrophils were increased following IL-1; however, this increase was reduced following IM. IL-1 marrow-derived CFU-GM, CFU-E, BFU-E, and CFU-Meg were below controls. In contrast, splenic CFU-GM and CFU-Meg were significantly elevated with increasing IL-1 concentrations. Erythroid progenitors were increased following low IL-1 concentrations and reduced in animals receiving IM, thus indicating a role for prostaglandins in the mechanism of IL-1 for influencing hematopoiesis. CFU-DG were increased, however, only when animals were pretreated with IL-1 and their cells implanted into normal hosts, not when normal cells were implanted into animals pretreated with IL-1, indicating a potential target cell effect rather than an indirect, factor-related response.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Heparin modulates the secretion of a major excreted protein-like molecule by vascular smooth muscle cells.

Previous work from our laboratory has shown that heparin specifically induces the release of a pair of proteins of approximately 35,000 and 37,000 Da into the culture medium of vascular smooth muscle cells (SMC). In this report, we demonstrate that the previously identified 37,000-Da smooth muscle protein is composed of two protein species with very similar molecular weights based on migration patterns in SDS-polyacrylamide gels. The larger molecular weight species in this doublet has a similar molecular weight and shares antigenic determinants with major excreted protein (MEP), a lysosomal proteinase previously shown to be secreted by normal and transformed fibroblasts and epidermal cells. Antisera to MEP precipitated the higher molecular weight band from the doublet; preimmune serum was not reactive with the smooth muscle protein. Exposure of smooth muscle cells to heparin resulted in decreased amounts of immunoprecipitable protein released into the medium. Thus, it now appears that three proteins in the 35,000-38,000 molecular weight range are modulated by heparin, and that the largest of the heparin-modulated vascular SMC proteins has a similar molecular weight and is immunologically related to MEP. The release of MEP-like protein from SMC is decreased by heparin, while the remaining two heparin-modulated proteins are increased in the presence of heparin.

Animals↗

Synergistic stimulation of neutrophils. Possible involvement of 5-hydroxy-6,8,11,14-eicosatetraenoate in superoxide release.

Neutrophils stimulated with optimal amounts of tumor-promoters that activate protein kinase C (e.g. mezerein, phorbol 12,13-dibutyrate) are known to release large quantities of superoxide: approximately 40-50 nmol O2-/min/10(7) cells. Previous studies have shown that treatment of neutrophils with the calcium ionophore A23187, or with 5-hydroxy-6,8,11,14-eicosatetraenonate (5-HETE), dramatically increased the ability of these cells to release O2- in response to suboptimal concentrations of the stimulants mentioned. In this manuscript, we provide data relevant to the basis of this augmentation of O2- release. The synergy with ionophore A23187 exhibited a partial requirement for extracellular Ca2+, whereas that with 5-HETE exhibited a near absolute requirement for that cation. Neutrophils stimulated with optimal amounts of tumor-promoters are known to exhibit a redistribution of protein kinase C activity from the soluble to a particulate fraction. A redistribution of kinase activity was not observed in cells stimulated synergistically. On the other hand, ionophore A23187 and 5-HETE increased the binding of a suboptimal amount of [3H] phorbol 12,13-dibutyrate to intact neutrophils by approximately 25 and 50%, respectively. Inhibitors of protein kinase C (i.e. sphingosine, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine) substantially blocked O-2 release from neutrophils stimulated either synergistically or with optimal levels of tumor-promoters. These data suggest a role for 5-HETE in modulating O-2 release by neutrophils and are discussed in relation to models of the interactions of protein kinase C with membranes.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Obliteration of mastoid cavities using bone pâté.

The authors report on a series of 24 patients in whom open cavity mastoidectomy cavities were obliterated with bone pâté. The aim of the procedure is to provide the patients with a dry ear which then also reduces the need for frequent out-patient attendances.

Cholesteatoma↗

A comparison of glyceryl trinitrate and labetalol as hypotensive agents in microsurgery of the middle ear.

Ninety-five patients undergoing microsurgery of the middle ear were anaesthetised using thiopentone, nitrous oxide, oxygen and halothane via a low flow circle system, with carbon-dioxide absorption. Systolic blood pressure was reduced to approximately 65 mmHg either by the use of increments of labetalol, or by an infusion of glyceryl trinitrate (GTN). The degree of haemostasis achieved was assessed by the surgeon who was unaware which hypotensive agent was being used. Both hypotensive agents produced good conditions in all patients. Thus GTN is a suitable agent to induce hypotension for this type of microsurgery.

Adolescent↗

Intrathoracic granular cell myoblastoma.

Intrathoracic granular cell myoblastoma (GCM) is most commonly a benign neoplasm of Schwann cell origin that occurs in the large airways, the segmental bronchi, and rarely in the mediastinum. The tumor predominantly affects blacks during the fourth decade, and shows no sex predilection. Tracheobronchial GCM most commonly produces cough and chest pain, with focal consolidation or atelectasis shown on chest x-ray films. Extraesophageal mediastinal GCM produces few or no symptoms, and should be included in the differential diagnosis of a posterior mediastinal mass. Computerized tomography is helpful in evaluating the extent of the lesion, defining its relationship to adjacent structures, and planning the therapeutic approach. The diagnosis of tracheobronchial GCM usually can be made by bronchoscopy and/or biopsy. Treatment remains controversial, but most studies suggest surgical or endoscopic resection as the treatment of choice. The prognosis of intrathoracic GCM is good but is dependent upon complete removal.

Adult↗

Spinach Leaf Chloroplast CO(2) and NO(2) Photoassimilations Do Not Compete for Photogenerated Reductant: Manipulation of Reductant Levels by Quantum Flux Density Titrations.

Potential competition between CO(2) and NO(2) (-) photoassimilation for photogenerated reductant (e.g. reduced ferredoxin and NADPH) was examined employing isolates of mesophyll cells and intact chloroplasts derived from mature ;source' spinach leaves. Variations in the magnitude of incident light energy were used to manipulate the supply of reductant in situ within chloroplasts. Leaf cell and plastid isolates were fed with saturating CO(2) and/or NO(2) (-) to produce the highest demand for reductant by CO(2) and/or NO(2) (-) assimilatory processes (enzymes). Even in the presence of CO(2) fixation, NO(2) (-) reduction in intact leaf cell isolates as well as plastid isolates was maximal at light energies as low as 50 to 200 microeinsteins per second per square meter. Simultaneously, 500 to 800 microeinsteins per second per square meter were required to support maximal CO(2) assimilation. Regardless of the magnitude of the incident light energy, CO(2) assimilation did not repress NO(2) (-) reduction, nor were these two processes mutually repressed. These observations have been interpreted to mean that reduced ferredoxin levels in situ in the plastids of mature source leaf mesophyll cells were adequate to supply the concurrent maximal demands exerted by enzymes associated with CO(2) as well as with inorganic nitrogen photoassimilation.

Journal Article↗

Haem synthesis from exogenous 5-aminolaevulinate in cultured chick-embryo hepatocytes. Effects of inducers of cytochromes P-450.

The effects of inducers of cytochrome P-450 on haem biosynthesis from 5-aminolaevulinate were examined by using cultured chick-embryo hepatocytes. Cultures treated with either 2-propyl-2-isopropylacetamide or 3-methylcholanthrene contained increased amounts of cytochrome P-450 and haem. After treatment for 3 h with 5-amino[4-14C]laevulinate, the relative amounts of radioactivity accumulating as haem corresponded to the relative amounts of total cellular haem, but not to increases in the amounts of cytochrome P-450. Treatment with 5-aminolaevulinate did not alter cellular haem or cytochrome P-450 concentrations in either control or drug-treated cultures. The mechanism of the enhanced accumulation of radioactivity in haem was investigated. Although 2-propyl-2-isopropylacetamide enhanced the uptake of 5-aminolaevulinate and increased the cellular concentration of porphobilinogen 1.5-fold, these changes did not account for the increases in haem radioactivity. The inducing drugs had no effect on the rates of degradation of radioactive haem, but appeared to enhance conversion of protoporphyrin into haem. This latter effect was shown by: (1) a decreased accumulation of protoporphyrin from 5-aminolaevulinate in cells treated with inducers, and (2) complete prevention of this decrease if the iron chelator desferrioxamine was present. We conclude that inducers of cytochrome P-450 may increase haem synthesis not only by increasing activity of 5-aminolaevulinate synthase, but also by increasing conversion of protoporphyrin into haem.

Allylisopropylacetamide↗

Interleukin-1 (IL-1) depresses cytochrome P450 levels and activities in mice.

Endotoxin depresses cytochrome P450 levels when injected into animals. The purpose of this study was to determine whether endotoxin itself, or monokine(s) released in response to endotoxin administration are responsible for this effect. Cytochrome P450 levels and drug metabolizing activities were measured in endotoxin resistant C3H/HeJ mice 24h after single intraperitoneal injections of either lipopolysaccharide (LPS), a semipurified murine monokine preparation containing interleukin-1 (IL-1), or murine recombinant IL-1. In endotoxin sensitive C3H/HeN mice, LPS (0.5 mg/Kg) decreased total cytochrome P450 levels, benzphetamine demethylase activities, and ethoxyresorufin-0-deethylase activities. This dose of LPS did not alter cytochrome P450 levels or activities in the C3H/HeJ mice. However, after injection of the semipurified monokine preparation or the recombinant IL-1, there were significant decreases in cytochrome P450 levels and activities similar to the decreases observed with LPS in the C3H/HeN mice. These findings suggest that the alterations in hepatic cytochrome P450 seen with endotoxin injection are mediated, at least in part, by IL-1.

Animals↗

A technique of tongue reconstruction following near-total glossectomy.

Total glossectomy is the most crippling of all oral cavity resections. Speech problems are unavoidable and post-operative aspiration is often severe. Two patients are presented who have had tongue reconstructions following near-total glossectomies. In each case a quilted, split-skin grafted pectoralis major muscle flap was used. The speech and swallowing results of these two patients are compared with two patients who underwent a hemiglossectomy and total glossectomy respectively without tongue reconstruction. Only if the suprahyoid and extrinsic tongue muscles are preserved can an active oral phase of swallowing and usable speech be regained by tongue reconstruction.

Carcinoma, Squamous Cell↗

Cell surface dynamics of neutrophils stimulated with phorbol esters or retinoids.

Neutrophils undergo rapid morphological changes as well as metabolic perturbations when stimulated with certain phorbol esters. Stimulated cells initially exhibit pronounced projections emanating from the cell bodies, followed by rounding of the cells, reduction in granule number, and the appearance of intracellular vesicles. We show these vesicles to be derived, at least in part, from the plasmalemma. The experimental approach involved labeling stimulated and unstimulated cells with native ferritin and cationized ferritin, along with the cytochemical localization of ecto-5'-nucleotidase. The labeling patterns of the vesicles indicate that these structures are involved in both phorbol ester-stimulated adsorptive and fluid-phase endocytosis. Neutrophils stimulated with 12-O-tetradecanoyl-phorbol-13-acetate (TPA) exhibit two distinct rates of superoxide release in which the second, prolonged level is approximately 50% of the initial rate. All-trans-retinal, which we have recently shown to stimulate O2- release but not granule exocytosis or cell vesiculation, induces a single prolonged rate of maximal O2- release. Neutrophils treated with both all-trans-retinal and TPA exhibit only a single sustained rate of maximal O2- release similar to that observed with all-trans-retinal alone. Moreover, treatment of cells with all-trans-retinal blocks the vesiculation of neutrophils induced by TPA in a dose-dependent manner. This observation provides a possible explanation for the differences in the kinetics of superoxide release.

Animals↗

Lack of pleiotropic compensation in extracellular protein production by hypoproducing variants of Staphylococcus simulans biovar staphylolyticus.

The changes in bacterial density, total extracellular protein and activities of three extracellular enzymes were monitored during growth of wild-type Staphylococcus simulans biovar staphylolyticus, a representative pleiotropic variant that produced decreased levels of the three extracellular enzymes, and a variant that produced only 5 of the 14 extracellular proteins secreted by the wild-type organism. Both variants produced less total extracellular protein than did the parental organism. SDS-PAGE of the proteins secreted by these hypoproducing variants showed that all of the extracellular proteins were produced in decreased amounts. No pleiotropic compensation in extracellular protein production was observed for these hypoproducing variants of S. simulans biovar staphylolyticus.

Bacterial Proteins↗

Adhesion of polymorphonuclear leukocytes to endothelium enhances the efficiency of detoxification of oxygen-free radicals.

Polymorphonuclear leukocytes can produce active oxygen species such as hydrogen peroxide and superoxide under various conditions. Because these substances can be toxic to cells, it is possible that the interaction between the circulating leukocytes and the blood vessel wall, either in normal circulation or during the acute inflammatory response, could damage the endothelial lining. Using an in vitro system of cultured endothelial cells and isolated polymorphonuclear leukocytes, we have measured the levels of detectable superoxide when neutrophils are attached to either endothelial monolayers or to plastic. Our results show that the levels of superoxide, on a per-cell basis, are lower when the neutrophils are attached to endothelium than when attached to plastic, even if the neutrophils are stimulated with phorbol myristate acetate. This is also reflected in data showing that no injury occurs to the endothelial cells, as measured by 51Cr release, under these same conditions. When endothelial cells are pretreated with an inhibitor of superoxide dismutase, diethyldithiocarbamate, the levels of superoxide detected are the same for neutrophils stimulated on plastic and those on the endothelial monolayer, suggesting that endothelial superoxide dismutase may remove a portion of the neutrophil-generated superoxide from the detection system. Further evidence for the role of endothelium in destroying superoxide is suggested by results that show that the level of detectable superoxide released from neutrophils attached to formalin-fixed endothelial monolayers is the same as that for neutrophils attached to plastic. It is important to note that with the inhibitor of superoxide dismutase present, the endothelial monolayers do not display enhanced 51Cr release under the conditions employed. When both endothelial catalase and glutathione reductase are inhibited, we detect increased 51Cr release from endothelial cells in response to stimulated neutrophils. Our results show that the endothelial cells are important in affecting the apparent reduction of toxic oxygen products derived from polymorphonuclear leukocytes attached to their surface.

Animals↗

Pectoralis major muscle flap in head and neck reconstruction.

Pectoralis major muscle flaps have rarely been used on their own for head and neck reconstruction. Some of the problems experienced with myocutaneous flaps can be avoided by the judicious use of muscle flaps. These include suture line separation, excessive bulk, hair growth from the flap, and alteration of breast position. In contrast to the pectoralis major myocutaneous flap, the pectoralis major muscle flap is light and pliable. When it is employed for reconstruction in the oral cavity, oropharynx, or hypopharynx, it can be covered by a "quilted" skin graft or used on its own. We believe that pectoralis major muscle flaps provide a valuable alternative to the more bulky myocutaneous flaps in head and neck reconstruction.

Aged↗