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Biomedical subjects

D J Smith

Publications and source records attributed to D J Smith.

At least 109 records · Page 6Linked to original sources
PubMed1999

Antibody to glucosyltransferase induced by synthetic peptides associated with catalytic regions of alpha-amylases.

We examined the immunogenicity and induction of inhibitory activity of 19-mer synthetic peptides which contained putative catalytic regions that were associated with the beta5 (EAW) and beta7 (HDS) strand elements of the suggested (beta,alpha)8 catalytic barrel domain of Streptococcus mutans glucosyltransferase (GTF). Both peptides readily induced serum immunoglobulin G (IgG) and salivary IgA antipeptide activity which was reactive both with the inciting peptide and with intact S. mutans GTF. Antisera to each peptide construct also inhibited the ability of S. mutans GTF to synthesize glucan. These observations support the existence of catalytic subdomains containing glutamate and tryptophan (EAW) or aspartate and histidine (HDS) residues, each of which have been suggested to be involved with the catalytic activity of GTF. Furthermore, the epitopes defined in these sequences have significant immunogenicity and can induce immune responses which interfere with GTF-mediated glucan synthesis.

Amino Acid Sequence↗
PubMed1999

Supraspinal neurotensin-induced antianalgesia in mice is mediated by spinal cholecystokinin.

Intracerebral injection of neurotensin into specific brain loci in rats produces hyperalgesia due to the release of cholecystokinin (CCK) in the spinal cord. The present purpose was to show in another species that neurotensin can antagonize the antinociceptive action of morphine through the spinal CCK mechanism in mice. Neurotensin given intracerebroventricularly (i.c.v.) at doses higher than 100 ng produced antinociception in the tail flick test. However, at lower doses between 1 pg to 25 ng, neurotensin antagonized the antinociceptive action of morphine given intrathecally (i.t.), thus demonstrating the antianalgesic activity of neurotensin. The rightward shift in the morphine dose-response curve produced by i.c.v. neurotensin was eliminated by an i.t. pretreatment with CCK8 antibody (5 microl of antiserum solution diluted 1:1000). I.t. administration of lorglumide, a CCK(A)-receptor antagonist (10-1000 ng), and PD135,158, a CCK(B)-receptor antagonist (250-500 ng), also eliminated the antianalgesic action of neurotensin. Thus, the mechanism of the antianalgesic action of neurotensin given i.c.v. involved spinal CCK. This mode of action is similar to that for the antianalgesic action of supraspinal pentobarbital which also involves spinal CCK.

Analgesics, Opioid↗
PubMed1999

Effects of ractopamine HCl stereoisomers on growth, nitrogen retention, and carcass composition in rats.

The objectives of this study were to determine the effects of ractopamine HCl (RAC) stereoisomers (RR, RS, SR, and SS) on performance, carcass composition, and nitrogen retention in growing female rats. Forty-eight rats (eight rats/treatment) were treated with 0 or 320 microg/d of RAC or with 80 microg/d of the RR, RS, SR, or SS stereoisomers of ractopamine. Rats had free access to feed and water before and during the experiment. Ractopamine and stereoisomers were delivered via i.p. implanted osmotic pumps for 14 d, and rats were then slaughtered. Control rats were fitted with osmotic pumps containing saline. Ractopamine increased (P < .05) feed intake (d 1 to 6); body weight; carcass CP; and intake, apparent absorption, retention, and retained:intake ratio of CP on d 1 to 6 of the study. Ractopamine decreased (P < .05) carcass lipid and visceral lipid. Rats dosed with the RR stereoisomer responded similarly to rats dosed with RAC, except for carcass lipid. Carcass lipid was decreased (P < .01) by RAC relative to controls, but it was not different from controls in rats treated with the RR isomer. Compared with controls, BW, carcass CP, and CP retention were increased by the RR stereoisomer, and visceral lipid was decreased. The RS isomer also decreased visceral lipid (P < .10), but variables measured in rats dosed with the RS, SR, and SS isomers generally did not differ from controls. Results of this study indicate that the RR isomer of RAC is responsible for a majority of the leanness-enhancing effects of RAC in rats.

Animals↗
PubMed1999

Bacterial colonization/infection and the surgical management of pressure ulcers.

The purpose of this paper is to review the current recommendations and guidelines for the care and treatment of pressure ulcers with specific reference to the control of infection within these wounds and surgical management. After reviewing the literature published between May 1993 and April 1998, it is our contention that no significant changes in the clinical management of this problem are warranted. This may signal the need for further study in this area. Recommendations for the optimal care of clean and infected pressure ulcers are included.

Bacterial Infections↗
PubMed1999

In vivo analysis of DNase I hypersensitive sites in the human CFTR gene.

BACKGROUND: The cystic fibrosis transmembrane conductance regulator gene (CFTR) shows a complex pattern of expression. The regulatory elements conferring tissue-specific and temporal regulation are thought to lie mainly outside the promoter region. Previously, we identified DNase I hypersensitive sites (DHS) that may contain regulatory elements associated with the CFTR gene at -79.5 and at -20.5 kb with respect to the ATG and at 10 kb into the first intron. MATERIALS AND METHODS: In order to evaluate these regulatory elements in vivo we examined these DHS in a human CFTR gene that was introduced on a yeast artificial chromosome (YAC) into transgenic mice. The 310 kb human CFTR YAC was shown to restore the pheno-type of CF-null mice and so is likely to contain most of the regulatory elements required for tissue-specific expression of CFTR. RESULTS: We found that the YAC does not include the -79.5 kb region. The DHS at -20.5 kb is present in the chromatin of most tissues of the transgenic mice, supporting its non-tissue-specific nature. The DHS in the first intron is present in a more restricted set of tissues in the mice, although its presence does not show complete concordance with CFTR expression. The intron I DHS may be important for the higher levels of expression found in human pancreatic ducts and in lung submucosal glands. CONCLUSION: These data support the in vivo importance of these regulatory elements.

Animals↗
PubMed1998

Isolation, structural characterization, and chromosomal mapping of the mouse vascular adhesion protein-1 gene and promoter.

Vascular adhesion protein-1 (VAP-1) is an endothelial cell adhesion molecule which mediates lymphocyte binding to endothelial cells. The cloning of a mouse VAP-1 (mVAP-1) cDNA revealed that mVAP-1 is a novel 110/220 kDa transmembrane molecule with significant identity to copper-containing amine oxidases. In this work the nucleotide sequence and primary structure of the mVAP-1 gene was determined and the promoter region was structurally characterized. The isolated approximately 14.4-kb mVAP-1 gene consists of 4 exons and 3 introns. Primer extension analysis and 5' rapid amplification of cDNA ends revealed multiple transcription initiation sites in different tissues suggesting that the mVAP-1 transcription is differently regulated in different tissues. Analysis of the sequence immediately upstream of the detected transcription initiation sites showed no canonical TATA or CCAAT elements, but putative regulatory elements were found close to the detected transcription start sites. The cloning of the mVAP-1 gene reveals the first insight into the genomic organization of murine amine oxidases and will, by targeted disruption of the gene, allow us to understand better the importance of VAP-1 in leukocyte trafficking and monoamine oxidase activity for the function of the immune system.

Amine Oxidase (Copper-Containing)↗
PubMed1998

Cloning of vascular adhesion protein 1 reveals a novel multifunctional adhesion molecule.

Vascular adhesion protein 1 (VAP-1) is a human endothelial sialoglycoprotein whose cell surface expression is induced under inflammatory conditions. It has been shown previously to participate in lymphocyte recirculation by mediating the binding of lymphocytes to peripheral lymph node vascular endothelial cells in an L-selectin-independent fashion. We report here that the VAP-1 cDNA encodes a type II transmembrane protein of 84.6 kD with a single transmembrane domain located at the NH2-terminal end of the molecule and six potential N-glycosylation sites in the extracellular domain. In vivo, the protein exists predominantly as a homodimer of 170-180 kD. Ax endothelial cells transfected with a VAP-1 cDNA express VAP-1 on their cell surface and bind lymphocytes, and the binding can be partially inhibited with anti-VAP-1 mAbs. VAP-1 has no similarity to any currently known adhesion molecules, but has significant identity to the copper-containing amine oxidase family and has a monoamine oxidase activity. We propose that VAP-1 is a novel type of adhesion molecule with dual function. With the appropriate glycosylation and in the correct inflammatory setting, its expression on the lumenal endothelial cell surface allows it to mediate lymphocyte adhesion and to function as an adhesion receptor involved in lymphocyte recirculation. Its primary function in other locations where it is expressed, such as smooth muscle, may depend on its inherent monoamine oxidase activity.

Amine Oxidase (Copper-Containing)↗
PubMed1998

Cloning and characterization of mouse vascular adhesion protein-1 reveals a novel molecule with enzymatic activity.

Human vascular adhesion protein-1 (VAP-1) is a sialylated endothelial cell adhesion molecule mediating the initial L-selectin-independent interactions between lymphocytes and endothelial cells in man. In this work we cloned and characterized mouse VAP-1 (mVAP-1) and produced an anti-mVAP-1 mAb against a recombinant mVAP-1 fusion protein. The isolated cDNA encodes a novel 84.5-kDa mouse molecule. The anti-mVAP-1 mAb stained high endothelial venules in peripheral lymph nodes, and smooth muscle cells and lamina propria vessels in gut. During immunoblotting, this anti-mVAP-1 mAb recognized a 110/220-kDa Ag, suggesting that mVAP-1 is a dimer. Since mVAP-1 has significant sequence identity to members of a family of enzymes called the copper-containing amine oxidases, we showed that mVAP-1 possesses monoamine oxidase activity. Thus, mVAP-1 is the first mouse membrane-bound amine oxidase identified at the molecular level. Based on the 83% identity between the isolated cDNA and human VAP-1 cDNA, the expression pattern, the molecular mass, and the enzyme activity against monoamines, the cloned molecule represents a mouse homologue of human VAP-1. Cloning of mVAP-1 provides a valuable tool for in vivo studies of the significance of VAP-1 for lymphocyte-endothelial cell interactions and of the possible relationship between leukocyte adhesion and amine oxidase activity.

Amine Oxidase (Copper-Containing)↗
PubMed1998

Ts1Cje, a partial trisomy 16 mouse model for Down syndrome, exhibits learning and behavioral abnormalities.

A mouse model for Down syndrome, Ts1Cje, has been developed. This model has made possible a step in the genetic dissection of the learning, behavioral, and neurological abnormalities associated with segmental trisomy for the region of mouse chromosome 16 homologous with the so-called "Down syndrome region" of human chromosome segment 21q22. Tests of learning in the Morris water maze and assessment of spontaneous locomotor activity reveal distinct learning and behavioral abnormalities, some of which are indicative of hippocampal dysfunction. The triplicated region in Ts1Cje, from Sod1 to Mx1, is smaller than that in Ts65Dn, another segmental trisomy 16 mouse, and the learning deficits in Ts1Cje are less severe than those in Ts65Dn. In addition, degeneration of basal forebrain cholinergic neurons, which was observed in Ts65Dn, was absent in Ts1Cje.

Animals↗
PubMed1998

Characterization and nitric oxide release studies of lipophilic 1-substituted diazen-1-ium-1,2-diolates.

Lipophilic esters of the naturally occurring polyamines putreanine and spermic acid were synthesized, characterized, and modified with nitric oxide to form the corresponding 1-substituted diazen-1-ium-1,2-diolates (previously known as NONOates). The resulting compounds were insoluble in water but were able to release nitric oxide (NO) when placed in phosphate buffered saline (PBS) at physiological pH. The two categories of compounds examined were putreanate esters of cholesterol or hexadecanol, and spermate diesters of cholesterol or hexadecanol. The putreanate NONOate esters of cholesterol and hexadecanol had NO release half-lives of 60 h and 81 h, respectively while the spermate NONOate diesters of cholesterol and hexadecanol and NO release half-lives of 23 days and 7.1 days, respectively. The presence of 5% Tween 20 increased the half-life of the cholesteryl putreanate NONOate to 104 h but only slightly increased the half-life of the hexadecyl putreanate NONOate to 89 h. The 1% presence of the pharmaceutical lung surfactant Survanta did not significantly alter the half-life of NO release for the cholesteryl putreanate NONOate or for the hexadecyl spermeate NONOate, but the surfactant did increase the amount of NO release by 46% and 67% respectively.

Amino Acids, Diamino↗
PubMed1998

Transdermal delivery of nitric oxide from diazeniumdiolates.

Adverse physiological effects can often interfere with the use of nitric oxide (NO) as a therapeutic agent, especially when it is systemically generated from prodrugs. NO which is generated and delivered site-specifically by transdermal donors may be useful in the treatment of parasitic, bacterial or viral skin infections without causing systemic side effects. Three diazeniumdiolates (formerly "NONOate"), including two water soluble compounds, (Z)-1-[N-2-aminoethyl)-N-(2-ammonioethyl)amino]-diazen-1- ium-1,2-diolate (DETA-NO) and (Z)-1-[N-(3-aminopropyl)-N-(3-ammoniopropyl)amino] diazen-1-ium-1,2-diolate (DPTA-NO), and one insoluble compound, DPTA-NO grafted to dextran microspheres (DPTA-NO-g-dextran) were used to transdermally deliver NO to rats. Dextran microspheres were obtained by simultaneously grafting DPTA-NO to dextran and cross-linking dextran with CNBr in an oil-water emulsion. Suspended in hydrogel, DETA-NO, DPTA-NO, and DPTA-NO-g-dextran were applied three times to depilated rats at 4 day intervals. Results show that metabolic urinary nitrate levels increase with time (24-48 h), reach a maximum, and return to baseline by the fourth day. DPTA-NO applications produced an average maximum nitrate level of 94.2 mumol/day +/- 34.2 mumol S.D. compared to the average maximum nitrate level of 39.8 mumol/day +/- 8.6 mumol S.D. obtained from treatment with DETA-NO. These results suggest that DPTA-NO delivered NO more efficiently than DETA-NO. When DPTA-NO-g-dextran microspheres were used as the NO donor, results comparable to DPTA-NO were observed with an average maximum nitrate level of 14.9 mumol/day +/- 3.0 mumol S.D. These and other conclusive data indicate that, via these diazeniumdiolates, NO can be delivered effectively through rat skin.

Administration, Cutaneous↗
PubMed1998

Using lazy evaluation to simulate realistic-size repertoires in models of the immune system.

We describe a method of implementing efficient computer simulations of immune systems that have a large number of unique B- and/or T-cell clones. The method uses an implementation technique called lazy evaluation to create the illusion that all clones are being simulated, while only actually simulating a much smaller number of clones that can respond to the antigens in the simulation. The method is effective because only 0.001-0.01% of clones can typically be stimulated by an antigen, and because many simulations involve only a small number of distinct antigens. A lazy simulation of a realistic number of clones and 10 distinct antigens is 1000 times faster and 10,000 times smaller than a conventional simulation--making simulations of immune systems with realistic-size repertoires computationally tractable.

Algorithms↗
PubMed1998

Effects of glutathione depletion on oxidant-induced endothelial cell injury.

Ischemia-reperfusion produces edema in vivo by disrupting endothelial cell junctional integrity. A cultured rat pulmonary artery endothelial cell (RPAEC) model was used to analyze the effects of oxidants and ischemic plasma in vitro. RPAEC cultures were treated with ischemic human plasma from transverse rectus abdominis musculocutaneous (TRAM) flaps following mastectomy or with an equal quantity of nonischemic plasma taken peripherally. Endothelial cells treated with ischemic plasma rounded and formed gaps within 5 min, then ruffled and blebbed after 10 min. Cultures treated with human nonischemic plasma had no gross morphological changes. Additionally, cultures treated with human ischemic plasma demonstrated an increase in diffusion rate of 125I-albumin across monolayers while monolayers treated with human nonischemic plasma had no increase in diffusion rate. RPAEC monolayers were treated with malic acid diethyl ester (DEM) or L-buthionine-[S, R]-sulfoximine (BSO) to decrease cellular stores of glutathione before exposure to oxidant stress. Cultures depleted of cellular glutathione stores were significantly (P < 0.05) more susceptible to 50 microM H2O2 than controls, as determined by an increase in diffusion rate of 125I-albumin across monolayers. To determine if ischemic plasma effects were mediated by oxidants, cultures were depleted of glutathione by DEM or BSO pretreatment before exposure to plasma from the ischemic hind limbs of Sprague-Dawley rats. Glutathione-depleted RPAEC monolayers were significantly (P < 0.05) and substantially (2-3 X) more susceptible to the effects of ischemic plasma than were cultures with normal glutathione levels. Glutathione depletion had no effect on cultures treated with an equal amount of nonischemic plasma from sham-operated rats. These data strongly suggest that ischemic plasma in the absence of any cellular component are able to induce an oxidant injury in endothelial cells and thereby compromise junctional integrity.

Animals↗
PubMed1998

Antigen direction of specific T-cell clones into gingival tissues.

This study was performed to investigate T-cell traffic to periodontal tissues during infection with a periodontal pathogen Actinobacillus actinomycetemcomitans (Aa). Rowett rat T-cell clones, A3 (CD4+ CD8-, alpha beta TCR+, NKRP-1-, specific to Aa) and G2 (CD4- CD8-, alpha beta TCR+, NKRP-1+, which reacts to Aa, Gram-negative and -positive bacteria), both expressed the same prominent adhesion molecules (LFA-1, VLA-4) to the same extent. Binding of both T-cell clones to rat endothelial cells in vitro was blocked by antibody to VLA-4. Rowett rats were infected with Aa and infused with Aa-stimulated, isogenic T-clone lymphocytes that had been labelled in vitro with 125IUdR. Radioactivity associated with recovery of clone A3, but not G2, was significantly elevated in the gingivae of infected rats, suggesting migration to infected animals' gingival tissues. Migration of radioactive Aa-specific A3 clone cells traced by autoradiography reached a maximum at 24 hr (1.2% of total lymphocytes as radiolabelled cells in infected gingiva versus 0.6% in noninfected), indicating an apparent antigen-directed retention in infected rats' gingival tissues. The G2 clone was not retained in the gingival tissues (0.20% of total lymphocytes as radiolabelled cells in infected gingiva versus 0.26% in non-infected). However, the possibility of A3 retention directed by inflammation or tissue-selective homing could not be excluded. In further experiments, other adoptively transferred T-clone lymphocytes [clones G23 (Th1) and F13 (Th2)] with specificity for the 29,000 MW outer membrane protein of Aa with the same prominent adhesion molecules could be recovered from rat gingivae previously challenged with this antigen. However, transferred T-clone lymphocytes [clone G26 (Th1)] with specificity for a different Aa antigen were not recovered. Therefore, the dynamics of cell entry into periodontal lesions vary for activated T lymphocytes with different antigenic specificities, indicating the significance of antigen in lymphocyte traffic to periodontal tissues.

Actinobacillus Infections↗
PubMed1998

Structural model of the catalytic domain of an enzyme with cell adhesion activity: human vascular adhesion protein-1 (HVAP-1) D4 domain is an amine oxidase.

Human vascular adhesion protein-1 (HVAP-1) is a multifunctional protein having at least two different cellular roles, functioning both as a lymphocyte-endothelial cell adhesion protein and as an enzyme with monoamine oxidase activity. HVAP-1 is a 180 kDa homodimeric glycoprotein consisting of a membrane-spanning domain and three predicted extracellular copper-containing amine oxidase domains. In HVAP-1 the extracellular domains are composed of a large domain D4, containing the active site and forming the interface of the dimer, while the smaller D2 and D3 domains surround the D4 dimer near the entrance to the active site. The structural model of the catalytic D4 domain of HVAP-1 reveals that all components necessary for enzymatic monoamine oxidase activity are indeed present within the HVAP-1 and pinpoints residues that may be key to substrate entry through a channel to the active site and residues likely to be involved in substrate specificity as well as structural features critical to dimer formation. Proper glycosylation is required for the cell adhesion function of HVAP-1 and the predicted location of the sugar units at the solvent-exposed surface suits this function well.

Amine Oxidase (Copper-Containing)↗