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Biomedical subjects

S A Thompson

Publications and source records attributed to S A Thompson.

At least 127 records · Page 7Linked to original sources
PubMed1991

Basic fibroblast growth factor is a beta-rich protein.

The conformation of the 153-residue form of human basic fibroblast growth factor (bFGF) was studied with circular dichroism (CD) and sequence prediction methods. The far-UV CD spectrum with a minimum at 202 nm resembled that of an unordered polypeptide/protein or a protein rich in distorted antiparallel beta-sheets. Analysis of the CD spectrum by the least-squares method of Chang et al. (1978) and the CONTIN program of Provencher and Glöckner (1981) suggested that about one half of the molecule consisted of beta-sheet and there was no alpha-helix. These estimates agreed with the prediction by the sequence method of Garnier et al. (1978) using decision constants based on CD results. bFGF had an unusual CD band at 187 nm, which disappeared upon ionization of Tyr side chains at pH 11.7. It also had another unusual property of irreversibly converting the CD spectrum to a helix-like one with a double minimum at 205 and 215 and a maximum at 189 nm upon heating the solution to above 55 degrees C. The helicity was also enhanced in trifluoroethanol and in sodium dodecyl sulfate. The mutant bFGF in which cysteines 76 and 94 were replaced by serine residues had essentially the same properties as the wild-type.

Amino Acid Sequence↗
PubMed1991

Endothelial cell viability in the rat aortic wall.

Aortic allografts may offer advantages over prosthetic materials for aortic valve replacement or reconstruction with a valved aortic conduit. Cellular viability may partly determine long-term allograft performance. To evaluate endothelial cell viability in a rat model, valved aortic conduits were subjected to collagenase digestion. The resulting endothelial cell suspension was labeled with Griffonia simplicifolia agglutinin-fluorescein isothiocyanate (GSA-FITC), a marker specific for vascular endothelial cells of the rat. The cells were then incubated with propidium iodide, which is excluded by viable cells. Flow cytometry evaluated endothelial cell viability by determination of percentage of GSA-FITC-positive cells that were negative for propidium iodide. Aortas were studied immediately after harvest or after storage at 4 degrees C in a nutrient medium for 3 to 21 days. Percentage of viable endothelial cells showed a progressive decline with increasing duration of storage. These results demonstrate flow cytometric measurement of endothelial cell viability, a factor of possible importance in assessing allograft storage methods, and show that endothelial viability declines with prolonged storage at 4 degrees C in a nutrient medium.

Animals↗
PubMed1991

Immunogenicity, antigenicity, and endothelial viability of aortic valves preserved at 4 degrees C in a nutrient medium.

In patients undergoing aortic valve replacement, allograft valves stored at 4 degrees C in a nutrient medium have been associated with excellent immediate and long-term results. The effects of this method of prolonged storage on the antigenic, immunological and cellular characteristics of these grafts are incompletely understood. This study was designed to study these phenomena in rat aortic valves subjected to antibiotic sterilization and stored for up to 3 weeks in RPMI containing 10% fetal calf serum. Selected valves from Brown Norway rats were implanted heterotopically into the abdominal aorta of Lewis rats. Other valves were studied prior to transplantation. Antigenicity was determined by immunocytochemical staining using monoclonal mouse antibodies directed at Class I and Class II rat antigens. Immunogenicity was determined by duration of second-set skin graft survival following heterotopic aortic valve implant. Endothelial cell viability was determined by flow cytometric analysis of endothelial cells harvested from aortic valve allografts by collagenase digestion. Only fresh valves and valves stored for 1 day were positive for Class I antigens; no valves were positive for Class II antigens. Duration of skin graft survival was prolonged with greater duration of storage, but grafts remained immunogenic after 21 days of storage. Endothelial cell viability declined from 95% in the fresh valves to 64% after 21 days of storage. With prolonged duration of allograft valve storage at 4 degrees C, there is an attenuation of antigenicity, immunogenicity, and endothelial cell viability. Loss of endothelial cells may contribute to the changes in immunological responses to the valve allografts. The expression of antigens on the endothelial surface is not a reliable predictor of immunological response.

Animals↗
PubMed1990

Molecular cloning and deletion of the gene encoding aspergillopepsin A from Aspergillus awamori.

We have cloned genomic pepA sequences encoding the aspartic proteinase aspergillopepsin A (PEPA) from Aspergillus awamori using a synthetic oligodeoxyribonucleotide probe. Nucleotide sequence data from the pepA gene revealed that it is composed of four exons of 320, 278, 249, and 338 bp. Three introns which interrupt the coding sequence are 51, 52, and 59 bp in length. Directly downstream from the putative start codon lies a sequence encoding 69 amino acids (aa) which are not present in mature PEPA. Based on similarities to other aspartic proteinases, this region may represent a 20-aa signal peptide followed by a 49-aa propeptide that is rich in basic aa residues. Northern blots of total cellular RNA extracted from A. awamori cells indicate that pepA is transcribed as a single 1.4-kb mRNA. Mutants of A. awamori lacking the pepA structural gene were derived by the following gene replacement strategy. First, we constructed a plasmid in which a 2.4-kb SalI fragment containing the entire pepA coding region was deleted from a 9-kb Eco RI genomic DNA clone and replaced by a synthetic DNA polylinker. Second, a selectable argB gene was inserted into the polylinker. Third, the EcoRI fragment which contained the argB marker flanked by pepA sequences was excised from the plasmid and used to transform an argB auxotroph of A. awamori. From 16-40% of the resulting prototrophic transformants were found to have a PEPA-deficient phenotype when screened with an immunoassay using antibodies specific for PEPA. Southern hybridization experiments confirmed that these mutants resulted from a gene replacement event at the pepA locus.

Amino Acid Sequence↗
PubMed1990

Cloning, characterization, and expression of two alpha-amylase genes from Aspergillus niger var. awamori.

Using synthetic oligonucleotide probes, we cloned genomic DNA sequences encoding an alpha-amylase gene from Aspergillus niger var. awamori (A. awamori) on a 5.8 kb EcoRI fragment. Hybridization experiments, using a portion of this cloned fragment to probe DNA from A. awamori, suggested the presence of two alpha-amylase gene copies which were subsequently cloned as 7 kb (designated as amyA) and 4 kb (amyB) HindIII fragments. DNA sequence analysis of the amyA and amyB genes revealed the following: (1) Both genes are arranged as nine exons and eight introns; (2) The nucleotide sequences of amyA and amyB are identical throughout all but the last few nucleotides of their respective coding regions; (3) The amyA and amyB genes from A. awamori share extensive homology (greater than or equal to 98% identity) with the genes encoding Taka-amylase from A. oryzae. In order to test whether both amyA and amyB were functional in the genome, we constructed vectors containing gene fusions of either amyA and amyB to bovine prochymosin cDNA and used these vectors to transform A. awamori. Transformants which contained either the amyA- or amyB-prochymosin gene fusions produced extracellular chymosin, suggesting that both genes are functional.

Amino Acid Sequence↗
PubMed1990

Effect of storage at 4 degrees C in a nutrient medium on antigenic properties of rat aortic valve allografts.

The effects of prolonged storage at 4 degrees C in nutrient medium on the antigenic properties of aortic valve allografts are unknown. Lewis rats received heterotopic aortic valve allografts from Brown Norway donors. Valves were transplanted immediately after harvest (fresh), or after antibiotic sterilization and storage in a nutrient medium at 4 degrees C for 3, 7, 14, or 21 days. Additional rats underwent sham laparotomy (sham). All recipient rats received Brown Norway skin grafts 3 weeks after valve transplant or sham procedure. Time to skin graft rejection for all groups was as follows: fresh (n = 10), 4.5 +/- 0.9 days; sham (n = 10), 7.1 +/- 0.3 days; 3-day (n = 10), 4.9 +/- 0.3 days; 7-day (n = 10), 5.2 +/- 0.4 days; 14-day (n = 10), 6.1 +/- 0.7 days; and 21-day (n = 10), 6.0 +/- 0.6 days. Significant differences existed between the sham group and each of the transplanted groups. No significant differences existed between the fresh group and either the 3-day or 7-day groups. The difference between the fresh group and the 14-day group approached significance (0.05 less than p less than 0.10), and the difference between the fresh and 21-day groups was significant (p less than 0.05). Additional valves not used for transplantation were studied with scanning electron microscopy. The valves preserved in nutrient medium exhibited a progressive loss of endothelium as compared with fresh valves. Storage of aortic valve allografts in a nutrient medium at 4 degrees C is associated with a progressive attenuation of antigenic response.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

A pleiotropic iron-uptake mutant of Neisseria meningitidis lacks a 70-kilodalton iron-regulated protein.

We isolated an iron-uptake mutant of Neisseria meningitidis M986-NCV-1 that was severely limited in the ability to use several sources of iron in the form of Fe3+. This mutant, FAM11, grew poorly or not at all with human transferrin (TF) or lactoferrin (LF) as the sole iron source in a defined medium but grew as well as wild-type meningococci with hemin or hemoglobin. Uptake of 55Fe bound to TF, LF, dicitrate complexes, aerobactin, or nitrilotriacetate was reduced to 0 to 4% of the wild-type level. FAM11 did not produce an iron-repressible outer membrane protein (FeRP) of 70 kilodaltons (kDa) found in membranes of iron-stressed M986-NCV-1. Western blot (immunoblot) analysis using rabbit antiserum against this protein revealed that at least 17 of 18 meningococcal and 10 of 14 gonococcal strains produced an FeRP of ca. 70 kDa. The 70-kDa FeRP was shown to be surface exposed by radioimmunoprecipitation with human immune sera. These data suggest that the 70-kDa FeRP is somehow involved in Fe uptake from TF and LF. However, we were unable to transform the iron-uptake phenotype from FAM11 into wild-type meningococci to confirm this. Revertants of FAM11 that grew with TF and LF did not regain the ability to make the 70-kDa FeRP but also did not completely regain the Fe-uptake phenotype of M986-NCV-1.

Antigens, Bacterial↗
PubMed1988

Absence of the coronary sinus.

The rare absence of the coronary sinus is described in a 78 year old male cadaver. The veins of the posterior surface of the heart, including the marginal and small cardiac veins, drained into the right atrium through an ostium guarded by a valve. The great cardiac vein drained into the superior vena cava after passing upward and beneath the left coronary artery.

Aged↗
PubMed1988

Anomalous fascicle and high origin of latissimus dorsi compensating for absence of serratus anterior.

The unilateral absence of the right serratus anterior muscle was found in a 78 year old white male subject. In addition, on the same side, a high origin (5th thoracic level) of latissimus dorsi with an accessory muscle fascicle which covered the inferior angle of the scapula and fused with teres major and with the deep fascia of infraspinatus was also found. The structural arrangement suggests that the absence of serratus anterior was compensated for by the anomalous latissimus dorsi.

Aged↗
PubMed1988

Effects of xylazine on cecal mechanical activity and cecal blood flow in healthy horses.

Mechanical activity of the cecal body, lateral cecal arterial blood flow, carotid arterial pressure, and heart rate were measured in 6 conscious healthy horses 30 minutes before and for 120 minutes after IV administration of xylazine at dosages of 1.1 mg/kg of body weight, 0.55 mg/kg, and 0.275 mg/kg. Xylazine at a dosage of 1.1 mg/kg reduced the mean motility index (the product of the mean amplitude of contractions and the total duration of contractile activity divided by the recording time) of the circular and longitudinal muscle layers for the first, second, third, and fourth 30-minute periods after administration of xylazine. Xylazine at a dosage of 0.55 mg/kg reduced the motility index of the circular and longitudinal muscle layers for the first and second 30-minute periods after administration of xylazine. Xylazine at a dosage of 0.275 mg/kg reduced the motility index of the circular and longitudinal muscle layers for the first 30-minute period after administration of xylazine. Mean lateral cecal arterial blood flow was significantly (P less than 0.05) lower than the base-line value at 2 and 4 minutes after administration of all 3 xylazine dosages and at 8 minutes after administration of xylazine dosages of 1.1 mg/kg and 0.55 mg/kg. All dosages of xylazine caused transient hypertension and bradycardia, followed by hypotension.

Animals↗
PubMed1987

Noninvasive measurement of gastric emptying in obstetric patients.

An epigastric impedance technique was used to measure gastric emptying in nonpregnant women, women in the third trimester of pregnancy, and women within 60 min of delivery. The basis of the technique is the increase in epigastric impedance after the ingestion of a nonionic fluid. The subsequent decline in impedance is used as a measure of gastric emptying. The impedance epigastrograph used consisted of a four-electrode array (two source, two detector) that was applied over the epigastrium. The method was sufficiently sensitive to measure the effects of pregnancy, labor, and narcotics on gastric emptying. There was no difference in the rate of gastric emptying between pregnant and nonpregnant females. Labor appeared to cause a significant delay in gastric emptying; the mean +/- SEM time taken for the gastric volume to decrease to 50% (T0.5) was 7.2 +/- 0.6 min during pregnancy and 13.0 +/- 1.9 min in the immediate postpartum period. However, the principal factor associated with the delay in gastric emptying in the group studied within 60 min of delivery was probably the use of meperidine and promethazine, because there was a significant difference (P less than 0.05) between subjects who received meperidine and promethazine during labor and those in another group who had received either no analgesia or extradural analgesia, the T0.5 values for the two groups being 18.2 +/- 4.0 and 10.3 +/- 1.4 min, respectively.

Drinking↗
PubMed1987

Outcomes of twin gestations at the University of Colorado Health Sciences Center, 1973-1983.

A retrospective study was undertaken of 341 twin pregnancies over a ten-year period at the University of Colorado Health Sciences Center. The perinatal morbidity and mortality were higher than for singleton gestations, but no difference was found between the first and second twins. Bed rest was effective in prolonging gestation and decreasing perinatal mortality (P less than .05). Delivery of the second twin in noncephalic presentation was accomplished vaginally in 46.5%, with external version successful in five of six attempts. The interval between the birth of each twin did not affect outcome.

Bed Rest↗