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Biomedical subjects

S A Thompson

Publications and source records attributed to S A Thompson.

At least 109 records · Page 6Linked to original sources

Augmentation of oral tissues in rabbit using autogenous fat.

Patients with mild velopharyngeal incompetence (VPI) may have speech disorders, which are not sufficiently severe to warrant extensive surgical intervention, yet may not be amenable to correction by speech therapy alone. Augmentation of the posterior pharyngeal wall to aid in closure of the velopharyngeal sphincter may be beneficial in establishing better speech patterns, especially when combined with speech therapy. A variety of materials and techniques have been used in the past for this purpose. In this setting, autogenous fat may be transplanted without the risks incurred by augmentation with synthetic materials and involves very little donor site morbidity. The literature is somewhat contradictory, however, regarding the stability of the augmentation achieved using autogenous fat and there are no histologic studies describing the fate of fat injected into tissues of the oral cavity. Prior to introduction of this technique into clinical practice, this study was designed to investigate the fate of autogenous fat injected submucosally in the oropharyngeal region. Autogenous fat was injected into the anterior soft palate using the rabbit as a model. Histologic and gross inspections were performed at 2 days, 1, 2, and 4 weeks after injections. At the end of 4 weeks, at least 50% of the injection sites had visible evidence of augmentation, and 90% had histologic evidence of submucosal fat. In some instances most of the fat was resorbed; however, there were no instances of clinical infection or necrosis of the injection site. We conclude that submucosal injection of autogenous fat is a feasible alternative to using synthetic or other biologic materials for augmentation in the oral cavity.

Adipose Tissue↗

Divergence of genetic sequences for the vacuolating cytotoxin among Helicobacter pylori strains.

Approximately 50% of Helicobacter pylori isolates produce a cytotoxin in vitro that induces vacuolation of eukaryotic cells. Screening a lambda ZapII library of H. pylori 60190 chromosomal fragments permitted the identification of a 3864-base pair (bp) open reading frame (vacA) that encoded the vacuolating cytotoxin, and a > or = 567-bp upstream gene that was homologous to Escherichia coli cysteinyl-tRNA synthetase. The sequence data suggest that a 33-amino-acid leader sequence and a C-terminal peptide are cleaved from a 139-kDa protoxin to yield the mature 87-kDa cytotoxin. The vacA gene product contains a C-terminal motif that is present in several other bacterial proteins that undergo C-terminal cleavage, including IgA proteases of Haemophilus influenzae and Neisseria gonorrhoeae. Isogenic H. pylori mutants with insertional mutation of the vacA gene lacked vacuolating cytotoxin activity and failed to produce the 87-kDa protein. Southern analysis of naturally occurring tox-H. pylori strains with vacA probes indicated the presence of hybridizing bands, but both Southern analysis and polymerase chain reaction studies suggested that the vacA sequences of tox- strains differed from those of tox+ strains. Sequence analysis of a 1541-bp region of polymerase chain reaction-amplified vacA from tox- strain 87-203 indicated 64.8% amino acid identity with the corresponding region from tox+ strain 60190. Thus, sequence divergence in vacA genes may explain the lack of functionally active cytotoxin production by some H. pylori isolates.

Amino Acid Sequence↗

Characterization of sequences within heparin-binding EGF-like growth factor that mediate interaction with heparin.

Heparin-binding (HB) epidermal growth factor (EGF)-like growth factor (HB-EGF), a member of the EGF protein family, is a potent mitogen for fibroblasts, smooth muscle cells, and keratinocytes that was initially identified as a secreted product of macrophage-like cells. HB-EGF and EGF appear to act on target cells utilizing the same receptor, but HB-EGF is distinguishable from EGF by its strong affinity for heparin. To facilitate studies of structure-function relationships in HB-EGF, a bacterial recombinant expression system was established that produced biologically active HB-EGF with the expected disulfide bonding pattern. Mutagenesis and protease digestion studies of the recombinant HB-EGF, coupled with heparin-binding analyses of synthetic peptides, indicated that the sequences within HB-EGF mediating its interaction with heparin are located primarily in a stretch of 21 amino acids characterized by a high content of lysine and arginine residues. Most of this heparin-binding domain lies in an amino-terminal region of HB-EGF that has no counterpart in EGF, but a portion of the 21-residue sequence extends into the EGF-like region of HB-EGF. In addition, the mutagenesis and synthetic peptide studies indicated that sequences in HB-EGF lying outside of the 21-residue stretch can also influence the interaction with heparin. Finally, a synthetic peptide derived from the 21-residue stretch was found to compete with HB-EGF for binding to Chinese hamster ovary cells, suggesting that the heparin-binding sequences in HB-EGF may also mediate the interaction of this factor with cell surface heparan sulfate proteoglycan.

Amino Acid Sequence↗

Effects of argon beam coagulation on split thickness skin graft survival in the swine model.

Argon beam coagulation (ABC) uses argon gas to evacuate blood in an operative field and simultaneously to transport a coagulating electric current to bleeding sites. This allows a 'no touch' method of coagulation which is faster, more precise, and less destructive than conventional electrocautery. The technique has been used to reduce blood loss in liver surgery, trauma surgery and neck dissections. In this study, the effect of ABC treatment of the recipient graft beds on the survival of split thickness skin grafts was assessed in pigs, testing the hypothesis that ABC treatment of the recipient bed would have no adverse effect on skin graft survival. Nine 20 kg mixed breed pigs had split thickness skin grafts raised on each side of the paraspinous area. The graft beds on one side were prepared using ABC and the contralateral side served as controls. The percentage of graft survival on the ABC treated sides was compared to control graft survival on the other side at 12 days postsurgery. Our results confirm the hypothesis that treatment of the recipient bed with ABC does not adversely affect the survival of split thickness skin grafts.

Animals↗

The effects of tretinoin on random skin flap survival in the swine model.

Tretinoin is currently marketed for topical treatment of acne vulgaris and has also been used in the rejuvenation of aging skin. After topical application, it has been shown to stimulate mitotic activity and increase vascularity in skin. In this study a porcine model was used to test the hypothesis that presurgical treatment with topical tretinoin would increase the surviving area of random full-thickness skin flaps. Four hybrid barrows, 40 to 60 lb, were treated with tretinoin and 4 similar animals were treated with placebo (carrier vehicle) for 2 weeks before raising four dorsally based full-thickness skin flaps (4 x 12 cm) on each animal. Biopsies were taken from tretinoin-treated, placebo-treated, and untreated skin at the time the flaps were initially raised and 1 week later before killing the animals. All tissue was processed for light and electron microscopy. One week after surgery, the pigs were killed. Photographs were taken at the termination of the experiment and the negatives were digitized and analyzed using a high-speed graphics workstation supported by SGITrace software. The percentage of skin flap survival was determined using this method of image analysis. Using repeated-measures analysis of variance, there was no significant difference in the mean flap survival between the tretinoin (mean = 46.75; SEM = 7.05) and placebo (mean = 65.80; SEM = 7.05) treated groups at the 0.05 significance level. Pretreatment with tretinoin did not enhance skin flap survival under the conditions of this study. Rationale for this finding and possible modifications of future studies are discussed.

Administration, Topical↗

Complication outcomes based on preoperative admission and length of stay for primary palatoplasty and cleft lip/palate revision in children aged 1 to 6 years.

With increasing focus on outcome studies, there is continued need for data about whether same-day admission and reduced hospital stay have adverse effects on surgical treatment, including that for cleft lip and palate. In this study, medical records were inspected for all cleft lip and palate patients, aged 1 to 6 years, who had primary palatoplasty or cleft lip/palate revision in this treatment center between 1978 and 1992 (N = 329). Length of stay for 251 (96.5%) of the 260 subjects admitted the day before surgery was from 4 to 7 days; 9 remained in the hospital longer than 8 days. Length of stay for 67 (97.1%) of 69 patients admitted the day of surgery was from 2 to 3 days; 2 were in the hospital for 7 days, and none for 8 or more days. Thirty-seven instances of surgical complications were reported for the 260 patients admitted the day before surgery (14.2%). Twelve complications (17.4%) were recorded for the 69 patients admitted the day of surgery. There was no significant difference in the number of complications between the two groups of patients (Fisher's exact test, p = 0.5682). There was no significant difference in the types of complications observed between the two groups (Fisher's exact test). Surgery was performed at age 1 year for 61 of the 69 patients admitted on the day of surgery (88.4%). The mean age of this group was significantly younger than that of patients operated on earlier than 1989 and admitted on the day before surgery (Wilcoxon's test, p = .0001, Z = 4.48).(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗

In utero cleft palate repair in the ovine model.

Cleft lip and palate defects assume many forms from mild to severe, but all may be associated with abnormal craniofacial development. Even the most expert and sophisticated methods of surgical repair are followed by scar contraction and fibrosis, which result in skeletal defects, dental abnormalities, cosmetic disfigurement, and speech impairment. Recent clinical and experimental observations that fetal cutaneous wounds heal without scarring are of great potential interest in the management of cleft lip and palate. The objective of this study was to investigate the effect of prenatal repair of iatrogenically produced cleft palate on scar formation in the fetal lamb model. Ten ewes were operated on ranging in gestation from 70 to 133 days. Fifteen lambs were studied (nine cleft palates produced and repaired in utero; one cleft produced in utero and not repaired, four normal, unoperated palates; and one cleft palate produced and repaired 1 week postnatally). The lambs were delivered normally at 145 to 147 days gestation and maintained with the ewe until 1 month of age. The lambs were euthanized, and the surgical area of the palates studied grossly and histologically. Animals operated at 112 days or later in gestation exhibited scars both clinically and histologically. The animals that had cleft palate produced and repaired at 70 days gestation did not have a visible palatal scar at 1 month of age. Histologically, there was evidence of minimal scarring without disruption of normal architecture. Studies are underway to determine the impact of reduced scarring on craniofacial growth after palatal repair during mid gestation in the ovine model.

Animals↗

Isolation and characterization of the Aspergillus oryzae gene encoding aspergillopepsin O.

We have cloned and determined the nucleotide sequence of a genomic DNA segment from Aspergillus oryzae which contains pepO, the gene encoding the aspartic proteinase, aspergillopepsin O (PEPO). The organization of pepO is strikingly similar to that of pepA from A. niger var. awamori (previously called A. awamori) in that both are composed of four exons and three introns with virtually identical lengths, and the positions of the introns are exactly conserved. From the deduced amino acid (aa) sequence, it appears that PEPO, like other fungal aspartic proteinases, is synthesized as a zymogen containing a putative N-terminal prepro-region of 77 aa followed by a mature protein of 327 aa. Southern blotting experiments suggest that a single copy of pepO exists in the A. oryzae genome.

Amino Acid Sequence↗

Cloning and nucleotide sequence of frpC, a second gene from Neisseria meningitidis encoding a protein similar to RTX cytotoxins.

Neisseria meningitidis FAM20 has recently been shown to produce two Fe-regulated proteins (FrpA and FrpC) related to the RTX family of cytotoxins. Here we report the cloning and DNA sequence of the locus containing the gene encoding the larger meningococcal RTX protein FrpC. FrpC was highly similar to FrpA throughout much of the predicted protein, with two main differences. Whereas the FrpA protein had 13 copies of the nine-amino-acid repeat units typical of RTX proteins, FrpC had 43 copies. The additional copies in FrpC apparently arose from a threefold tandem amplification of a 600bp DNA fragment encoding the repeats. In addition, the frpC gene lacked good promoter consensus sequences. An open reading frame (ORF1) of unknown function was found immediately upstream of frpC, suggesting the possibility that frpC was cotranscribed with ORF1. A probable promoter was found 300bp upstream of ORF1, and it contained a Fur protein-binding sequence found in the promoters of Fe-regulated Escherichia coli genes. DNA upstream of the ORF1/frpC promoter was homologous to IS1016-like elements surrounding capsulation loci of strains of Haemophilus influenzae. A FrpC-like protein (reactive in immunoblots with monoclonal antibody 9D4; multiple reactive bands of about 200 to 120 kDa) was found in five out of eight meningococcal strains but only in one out of 14 other Neisseria, suggesting that FrpC may participate in the pathogenesis of meningococcal disease.

Amino Acid Sequence↗

The RTX cytotoxin-related FrpA protein of Neisseria meningitidis is secreted extracellularly by meningococci and by HlyBD+ Escherichia coli.

Neisseria meningitidis produces proteins (FrpA and FrpC) related to the RTX cytotoxin family. In meningococcal strain FAM20 these proteins were both localized in the outer membrane and secreted into the extracellular medium. An Escherichia coli strain with wild-type hemolysin secretion genes hlyB and hlyD and containing a cloned frpA gene secreted FrpA, whereas an isogenic hlyBD mutant strain did not. Low-stringency DNA hybridization revealed hlyBD-like sequences in N. meningitidis FAM20, suggesting that a similar RTX secretion system exists in meningococci. Structural features found at the C termini of other RTX proteins and thought to be important for their secretion were also found at the C terminus of FrpA. The secretion of FrpA from E. coli by heterologous RTX transport proteins further demonstrates the relation of the FrpA protein to RTX toxins.

Amino Acid Sequence↗

Neisseria meningitidis produces iron-regulated proteins related to the RTX family of exoproteins.

A monoclonal antibody (A4.85) which reacted with Fe-regulated proteins of Neisseria meningitidis, was used to isolate a lambda gt11 clone from N. meningitidis FAM20. Chromosomal fragments flanking the fragment expressing the A4.85 epitope were cloned, and their DNA sequences revealed a 3,345-bp open reading frame predicting a 122-kDa protein. This gene was named frpA (Fe-regulated protein). A computer similarity search of GenBank revealed high levels of similarity to members of the RTX family of cytotoxins, especially in a region of tandem 9-amino-acid repeats. These repeats are found in all members of the RTX family; similar repeats were present 13 times in the predicted FrpA protein. Antigenic relatedness between the meningococcal proteins and the RTX proteins was demonstrated by the reactivity of A4.85 with Escherichia coli hemolysin (HlyA) and Bordetella pertussis adenylate cyclase-hemolysin (CyaA). Similarly, FrpA was recognized by 9D4, a monoclonal antibody directed against B. pertussis CyaA. In addition to the frpA gene, a second gene (frpC) produced a larger RTX-related protein. The frpA and frpC loci were mutagenized in strain FAM20, resulting in the loss of RTX-related proteins. A 120-kDa protein was expressed from the reconstructed frpA gene in E. coli. The biological function of FrpA is unknown, but its similarity to other RTX toxins suggests that it may play an important role in the pathogenesis of meningococcal infection.

Amino Acid Sequence↗

Suitability of the iliac crest as a site for harvest of autogenous bone grafts.

Many donor sites have been advocated for obtaining cancellous bone to be used for grafting alveolar defects. Recently, some authors have suggested that the iliac crest site produced an unacceptably high degree of postoperative morbidity. Because of this morbidity, the use of other donor areas, e.g., rib, cranium, and mandible, are advocated. The iliac crest donor site for alveolar bone grafting has been common practice in our institution for years under the assumption that little or no postoperative morbidity occurred. To investigate and document the accuracy of our assumption, the charts of the most recent 50 consecutive patients receiving alveolar bone grafts, all of which used the iliac crest as a donor site, were reviewed. Postoperative pain, numbness, infection, paralysis, or long-term disability were catalogued from the charts. In addition, a questionnaire was sent to the parents and patients requesting their perceptions of these parameters. In the cases reviewed, the average length of skin incision was 4.2 cm. The average quantity of bone harvested was 4.1 cc. The usual hospitalization time was 2-3 days. No patients experienced serious or long-term complications at the surgical donor site. Immediate postoperative complications among the 50 patients studied were limited to one hematoma, one stitch abscess, one case of swelling with pain, one erythema, and one 4-day fever accompanied by slight serosanguineous drainage. Information from the questionnaire suggests that most patients returned to full activity within 4-6 weeks. No patient reported any long-term pain or disability.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Effects of magnetic resonance imaging fields on gold eyelid loads.

Magnetic resonance imaging is frequently used for visualization of soft tissues of the head and neck. It was the purpose of this study to determine whether gold eyelid weights, used for treatment of lagophthalmos in facial palsy, would be deflected at high magnetic strengths during imaging. Ex vivo study of six incrementally increasing weights at two magnetic strengths was performed. There was no deflection of any of the gold eyelid loads at either 0.5-tesla or 1.5-tesla magnetic strengths. We conclude that patients with gold eyelid weights are not at risk of damage from movement of the prostheses during magnetic resonance imaging examination.

Eyelids↗

The disulfide structure of bovine pituitary basic fibroblast growth factor.

Basic fibroblast growth factor has 4 cysteine residues in its amino acid sequence, two of which are perfectly conserved within the fibroblast growth factor family of proteins suggesting a disulfide bond at this position. Furthermore, thiol titration of bovine pituitary basic fibroblast growth factor (bFGF) indicates the presence of two free thiols, which is consistent with an intramolecular disulfide. Direct analysis of natural and recombinant fibroblast growth factor proteins have not confirmed the existence of such a disulfide. Instead, the two nonconserved cysteines of bFGF purified from bovine pituitaries are S-thiolated with glutathione. Inclusion of 75 mM N-ethylmaleimide during the homogenization of the pituitaries effectively blocks the S-thiolation, demonstrating that this modification is an artifact of the purification procedure. Analysis of the N-ethylmaleimide purified bovine pituitary bFGF suggests that the natural protein is in the correct redox state when all 4 cysteines are in the reduced form.

Animals↗

Effect of immunological differences on rat aortic valve allograft calcification.

Calcification may be a cause of allograft valve degeneration. To determine whether immunological differences between donor and recipient affect the degree of calcification that occurs, adult Lewis rats received aortic valve allografts transplanted heterotopically into the abdominal aorta. All valves were transplanted immediately after harvest. The valves were not exposed to antibiotics or albumin before insertion. Valve donors were of the Lewis (syngeneic), F344 (weakly allogeneic, RT1 compatible, non-RT1 incompatible), LBN F1 (moderately allogeneic, one haplotype identical, one haplotype incompatible at the RT1 and non-RT1 loci), and Brown Norway (strongly allogeneic, RT1 and non-RT1 incompatible) strains. Valves were harvested 3-12 weeks following transplantation. Scanning electron microscopy and energy dispersion x-ray microanalysis were performed on one leaflet of each valve to evaluate calcium content. Calcium content expressed in counts (mean +/- standard error) according to donor strain were: Lewis, 1642 +/- 233; F344, 4853 +/- 1412; LBN F1, 4714 +/- 823; and Brown Norway, 4358 +/- 835. Significant differences (p less than 0.05) existed between valves from Lewis donors and those from each other strain. No differences among the other strains were statistically significant. It is concluded that syngeneic valve allografts calcify less than allogeneic grafts. However, the degree of allogenicity did not influence the magnitude of calcification.

Analysis of Variance↗

Gonococcal transferrin-binding protein 1 is required for transferrin utilization and is homologous to TonB-dependent outer membrane receptors.

The pathogenic Neisseria species are capable of utilizing transferrin as their sole source of iron. A neisserial transferrin receptor has been identified and its characteristics defined; however, the biochemical identities of proteins which are required for transferrin receptor function have not yet been determined. We identified two iron-repressible transferrin-binding proteins in Neisseria gonorrhoeae, TBP1 and TBP2. Two approaches were taken to clone genes required for gonococcal transferrin receptor function. First, polyclonal antiserum raised against TBP1 was used to identify clones expressing TBP1 epitopes. Second, a wild-type gene copy was cloned that repaired the defect in a transferrin receptor function (trf) mutant. The clones obtained by these two approaches were shown to overlap by DNA sequencing. Transposon mutagenesis of both clones and recombination of mutagenized fragments into the gonococcal chromosome generated mutants that showed reduced binding of transferrin to whole cells and that were incapable of growth on transferrin. No TBP1 was produced in these mutants, but TBP2 expression was normal. The DNA sequence of the gene encoding gonococcal TBP1 (tbpA) predicted a protein sequence homologous to the Escherichia coli and Pseudomonas putida TonB-dependent outer membrane receptors. Thus, both the function and the predicted protein sequence of TBP1 were consistent with this protein serving as a transferrin receptor.

Amino Acid Sequence↗

Effects of calcium channel blockers on calcium uptake in rat aortic valve allografts.

BACKGROUND: The life span of human aortic valve allografts is finite, and many fail because of cusp rupture or calcification. Subcellular changes occurring in aortic valves in response to transplantation include the uptake of calcium. This study uses a heterotropic rat aortic valve transplant model to determine whether the calcium channel blockers diltiazem and verapamil might attenuate leaflet calcification. METHODS AND RESULTS: The 60 rats studied were divided into the following groups: 1) control: valves from normal, unoperated F1 generation of Lewis and Brown Norway cross (LBNF1) rats; 2) control: valves from syngeneic transplant combinations (Lewis/Lewis); 3) control: valves from allogeneic transplant combinations (LBNF1/Lewis, donor/recipient); 4) experimental: valves from allogeneic strain combinations treated with 30 mg/kg per day diltiazem; 5) experimental: valves from allogeneic strain combinations treated with 30 mg/kg per day verapamil. Drugs or saline (controls) were administered with osmotic pumps placed subcutaneously 2 days before transplantation. Animals were killed 3 weeks later, and the valves were harvested and prepared for calcium analysis. Energy-dispersive x-ray microanalysis was used to measure the calcium in a section of one leaflet from each valve studied. Paired t tests showed that allograft valves treated with diltiazem or verapamil contained significantly less calcium than allograft controls treated with saline (p < 0.001). When all five groups were subjected to one-way ANOVA, the valves in the allograft control group contained significantly more calcium than all other groups. All other groups were not different from each other. CONCLUSIONS: The calcium channel blockers verapamil and diltiazem were effective in preventing early calcification that occurs in aortic valves after transplantation. Thus, these agents might play a role in prolonging the life of human aortic valve allografts.

Analysis of Variance↗