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Biomedical subjects

L A Thomas

Publications and source records attributed to L A Thomas.

At least 37 records · Page 2Linked to original sources

Identification of a Brazil-nut allergen in transgenic soybeans.

BACKGROUND: The nutritional quality of soybeans (Glycine max) is compromised by a relative deficiency of methionine in the protein fraction of the seeds. To improve the nutritional quality, methionine-rich 2S albumin from the Brazil nut (Betholletia excelsa) has been introduced into transgenic soybeans. Since the Brazil nut is a known allergenic food, we assessed the allergenicity of the 2S albumin. METHODS: The ability of proteins in transgenic and non-transgenic soybeans, Brazil nuts, and purified 2S albumin to bind to IgE in serum from subjects allergic to Brazil nuts was determined by radioallergosorbent tests (4 subjects) and sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (9 subjects) with immunoblotting and autoradiography. Three subjects also underwent skin-prick testing with extracts of soybean, transgenic soybean, and Brazil nut. RESULTS: On radioallergosorbent testing of pooled serum from four subjects allergic to Brazil nuts, protein extracts of transgenic soybean inhibited binding of IgE to Brazil-nut proteins. On immunoblotting, serum IgE from eight of nine subjects bound to purified 2S albumin from the Brazil nut and the transgenic soybean. On skin-prick testing, three subjects had positive reactions to extracts of Brazil nut and transgenic soybean and negative reactions to soybean extract. CONCLUSIONS: The 2S albumin is probably a major Brazil-nut allergen, and the transgenic soybeans analyzed in this study contain this protein. Our study show that an allergen from a food known to be allergenic can be transferred into another food by genetic engineering.

2S Albumins, Plant↗

Virulence attributes of Escherichia coli isolated from dairy heifer feces.

Escherichia coli isolates from 1,305 (of 6,894) fecal samples collected during the 1991-1992 USDA, Animal and Plant Health Inspection Service, National Health Monitoring System, Diary Heifer Evaluation Project were tested for virulence attributes associated with human enterohaemorrhagic E. coli (EHEC) and the enterotoxin commonly associated with diarrhoea in newborn calves. Single, random isolates from each heifer were hybridized to probes derived from the 60 mDa EHEC plasmid (CVD 419), E. coli attaching and effacing gene (eae), Shiga-like toxin (slt) genes I and II, and E. coli heat-stable enterotoxin a (STaP). Seventy-seven of the 1305 isolates (5.9%) were slt-positive. Most (81.8%) slt-positive E. coli were also CVD 419 and eae-positive. Only 2 of the slt-positive E. coli isolates were STaP-positive.

Animals↗

Phenotypic and ribosomal RNA characterization of Arcobacter species isolated from porcine aborted fetuses.

Aerotolerant organisms resembling Campylobacter, now designated as Arcobacter, have been described from aborted farm animals and from cases of human enteritis worldwide. The goals of this study were 1) to attempt to recover Arcobacter spp. from cases of porcine abortion, 2) to characterize these isolates by phenotype and ribotype, and 3) to compare the usefulness of ribotype and phenotype patterns for identifying Arcobacter butzleri and the DNA hybridization groups 1A and 1B of A. cryaerophilus. Isolates of Arcobacter spp. from North Carolina and Iowa were recovered from porcine tissues. In Iowa, Arcobacter spp. were recovered from 43% (13/30) of porcine abortion cases evaluated. Isolations were made from placenta (44%), kidney (44%), and stomach contents (12%), which were the only tissues examined. The most reliable biochemical tests for A. butzleri included growth in 1% glycine and in 1.5% NaCl, weak catalase activity, and resistance to cadmium chloride. Arcobacter cryaerophilus strains were characterized by strong catalase activity and sensitivity to cadmium chloride. The DNA hybridization groups 1A and 1B of A. cryaerophilus could not be distinguished by biochemical tests. This represents the first description of A. cryaerophilus DNA group 1A in animals within the United States.

Abortion, Veterinary↗

Management factors related to Salmonella shedding by dairy heifers.

In a national survey of dairy operations with > or = 30 milking cows, fecal samples from 6861 preweaned dairy heifer calves on 1063 operations were tested for Salmonella. One hundred forty-five (2.1%) of the fecal samples from 79 (7.4%) of the farms tested positive for Salmonella. A multivariate logistic regression model revealed that medicated milk replacer and hay fed to dairy heifers from 24 h of age until weaning were associated with a reduced risk of Salmonella sheddings, as was calving in an individual animal area within a building.

Animal Feed↗

Treatment of commercial broiler chickens with a characterized culture of cecal bacteria to reduce salmonellae colonization.

The effect of treatment with a newly developed characterized continuous-flow (CCF) culture composed of 29 strains of cecal bacteria on salmonellae colonization was evaluated in commercially reared broiler chickens. Newly hatched chicks in three flocks were sprayed with CCF culture as they were placed in rearing houses and compared with untreated flocks on the same farm. Hatchery transport tray liners and rearing house feed, water, and little samples were cultured for the presence of salmonellae. Cecal samples were cultured after 3- and 6-wk growout. Skin-feather samples were cultured at 6-wk growout. Fifty-five percent of the transport tray liners were contaminated with salmonellae on the day of chick placement. At 3 wk, salmonellae serotypes present on the tray liners were widely distributed in the litter of the rearing houses. The results indicated that exposure to salmonellae occurred before culture treatment and continued in the rearing houses during the 6-wk growout period. Salmonellae cecal colonization was decreased (P < .05) in two of the treated flocks at 3 wk and diminished (P < .07) in the third treated flock compared with control flocks. At 6 wk, skin-feather contamination and cecal colonization were decreased (P < .05) in one of the treated flocks whereas no treatment effect occurred in two of the treated flocks compared with controls. The results clearly indicate the necessity of implementing integrated programs to control salmonellae in both the hatchery and rearing house environments. The CCF culture served to enhance salmonellae colonization resistance and may serve as a useful component of an integrated control program.

Animals↗

Single subunit chimeric integrins as mimics and inhibitors of endogenous integrin functions in receptor localization, cell spreading and migration, and matrix assembly.

The ability of single subunit chimeric receptors containing various integrin beta intracellular domains to mimic and/or inhibit endogenous integrin function was examined. Chimeric receptors consisting of the extracellular and transmembrane domains of the small subunit of the human interleukin-2 receptor connected to either the beta 1, beta 3, beta 3B, or beta 5 intracellular domain were transiently expressed in normal human fibroblasts. When expressed at relatively low levels, the beta 3 and beta 5 chimeras mimicked endogenous ligand-occupied integrins and, like the beta 1 chimera (LaFlamme, S. E., S. K. Akiyama, and K. M. Yamada. 1992. J. Cell Biol. 117:437), concentrated with endogenous integrins in focal adhesions and sites of fibronectin fibril formation. In contrast, the chimeric receptor containing the beta 3B intracellular domain (a beta 3 intracellular domain modified by alternative splicing) was expressed diffusely on the cell surface, indicating that alternative splicing can regulate integrin receptor distribution by an intracellular mechanism. Furthermore, when expressed at higher levels, the beta 1 and beta 3 chimeric receptors functioned as dominant negative mutants and inhibited endogenous integrin function in localization to fibronectin fibrils, fibronectin matrix assembly, cell spreading, and cell migration. The beta 5 chimera was a less effective inhibitor, and the beta 3B chimera and the reporter lacking an intracellular domain did not inhibit endogenous integrin function. Comparison of the relative levels of expression of the transfected beta 1 chimera and the endogenous beta 1 subunit indicated that in 10 to 15 h assays, the beta 1 chimera can inhibit cell spreading when expressed at levels approximately equal to the endogenous beta 1 subunit. Levels of chimeric receptor expression that inhibited cell spreading also inhibited cell migration, whereas lower levels were able to inhibit alpha 5 beta 1 localization to fibrils and matrix assembly. Our results indicate that single subunit chimeric integrins can mimic and/or inhibit endogenous integrin receptor function, presumably by interacting with cytoplasmic components critical for endogenous integrin function. Our results also demonstrate that beta intracellular domains, expressed in this context, display specificity in their abilities to mimic and inhibit endogenous integrin function. Furthermore, the approach that we have used permits the analysis of intracellular domain function in the processes of cell spreading, migration and extracellular matrix assembly independent of effects due to the rest of integrin dimers. This approach should prove valuable in the further analysis of integrin intracellular domain function in these and other integrin-mediated processes requiring the interaction of integrins with cytoplasmic components.

Amino Acid Sequence↗

Contact stimulation of cell migration.

Mass migrations of dense cell populations occur periodically during embryonic development. It is known that extracellular matrices, through which the cells migrate, facilitate locomotion. However, this does not explain how cells, such as neural crest, can migrate as a dense cohort of cells in essentially continuous contact with one another. We report here that unique behavioral characteristics of the migrating cells may contribute to cohesive migration. We used time-lapse video microscopy to analyze the migration of quail neural crest cells and of two crest derivatives, human melanoma cells and melanocytes. These cells migrated poorly, if at all, when isolated, but could be stimulated up to 200-fold to travel following contact with migrating cells. This phenomenon, which we have termed "contact-stimulated migration," appeared to activate and sustain migration of the mass of cells. Cells that became dissociated from the others ceased directional migration, thereby limiting aberrant cell dispersion. Fibroblasts were minimally responsive to this novel phenomenon, which may be crucial for major, mass cell migrations.

Animals↗

Relationship between colloid osmotic pressure and plasma protein concentration in cattle, horses, dogs, and cats.

The relationship between colloid osmotic pressure (COP) and protein concentration was investigated for purified proteins and plasma samples obtained from cattle, horses, dogs, and cats. At equivalent concentrations, bovine albumin exerted a COP that exceeded that of gamma-globulins by a mean factor of 4.4. Similar relationships between COP and protein were observed in the other species. Consequently, for a given total protein concentration, COP was dependent on the albumin/gamma-globulins ratio. A commonly used nomogram for estimating COP from protein concentration, the Landis-Pappenheimer equation, did not provide reliable results for plasma samples from these species.

Animals↗

Intraluminal radiation therapy per endoprosthesis: a case study.

Palliation of cholangeocarcinomas has been achieved by radiation delivered to the bile duct via endoprothesis. This case study supports and extends the works of others by describing the endoscopic techniques involved in the implantation of iridium seeds within a nasobiliary catheter. The case presented demonstrates endoscopic radiation treatment with no systemic radiation effects in a patient whose life extension was 19 months.

Adenocarcinoma↗

Atrial automatic tachycardia-reversion pacemakers: their economic viability and impact on quality-of-life.

UNLABELLED: Refractory supraventricular tachyarrhythmias may be both difficult and costly to control medically and can interfere with the patient's lifestyle. Newer treatment modalities are available for their management, and these require comprehensive assessment. We therefore compared costs and selective indices of patient benefit in a group of 17 patients in whom an atrial antitachycardia (Intermedics Intertach 262-12) pacemaker was placed for refractory supraventricular tachyarrhythmias. Prior medical therapy was compared to subsequent automatic antitachycardia pacemaker treatment. The total medical costs (admissions, emergency room visits, office visits, and medication costs) and the number of hospitalizations and medications were compared prior to implantation (F/U 69.3 +/- 61 months) and after implantation (F/U 15.3 +/- 7.8 months). A detailed quality-of-life questionnaire was also obtained 36.6 +/- 11 months after implantation. RESULTS: There were significant per patient differences in total cost before and after implantation: monthly costs were $505 +/- $833 before pacemaker implantation and $105 +/- $117 monthly afterward (P less than 0.005). Pacemaker implantation hospitalization costs were $19,063 +/- $8,362. Monthly medication costs averaged $46 before versus $15 after implantation (P less than 0.01). The number of medication types also differed with an average 5.5 medication types per patient before versus 1.2 after implantation (P less than 0.001). There were 8.6 yearly hospital admissions in the whole group before implantation, versus 4.7 admissions in the group per year thereafter. Patients demonstrated significant improvement in 80% of the quality-of-life parameters studied. CONCLUSION: Adjunctive atrial automatic tachycardia-reversion pacemaker therapy may be cost-competitive over time when compared to medical therapy alone in patients with refractory supraventricular tachyarrhythmias and appears to improve overall quality-of-life.

Costs and Cost Analysis↗

The influence of endocardial electrode fixation status on acute and chronic atrial stimulation threshold and atrial endocardial electrogram amplitude.

UNLABELLED: The endocardial atrial electrogram or "P wave amplitude" (PWA) and pacemaker atrial stimulation thresholds are important parameters determining correct pacing system function. Pacemaker lead fixation mechanism and lead age may negatively influence these parameters. Therefore, we compared acute and chronic PWA and atrial stimulation thresholds in 33 patients with permanent transvenous atrial screw-in leads, follow-up 647 days +/- 297 days; and in 31 patients with nonscrew leads, follow-up 855 days +/- 512 days (P = ns). RESULTS: The PWA differed between the two lead types acutely (1.97 mV +/- 0.8 mV for screw-in versus 2.48 mV +/- 1.1 mV for nonscrew-in leads; P less than 0.05), but not chronically (2.21 mV +/- 0.8 mV vs 2.2 mV +/- 1.2 mV; P = ns). Acute and chronic atrial pacing thresholds did not differ between groups. We also analyzed an early interim PWA in a subgroup of patients (mean 31 days after implantation). The nonscrew fixation group interim PWA was 1.76 mV +/- 0.9 mV versus 2.7 mV +/- 1.2 mV at implant (P less than 0.001). The screw-in lead interim PWA was 2.04 mV +/- 0.9 mV versus 1.97 mV +/- 0.7 mV at implant (P = ns). CONCLUSIONS: (1) A significantly higher endocardial PWA occurs at the time of lead implantation in nonscrew versus screw-in lead groups, but the chronic PWA does not differ between the two groups. (2) A transient but marked early (mean approximately 31 days) attenuation of the PWA occurs only with nonscrew-in leads. (3) Atrial threshold stimulation energies do not differ between the two lead groups acutely or during follow-up.

Aged↗

Variation in P wave amplitude immediately after pacemaker implantation: possible mechanism and implications for early programming.

The P wave amplitude (PWA) plays an important role in determining atrial sensing capabilities. To assess early PWA change, we compared the unipolar PWA in 43 patients at the time of atrial lead placement, measured by a pacing systems analyzer, to the unipolar PWA recorded at the end of pacemaker surgery, from telemetered atrial endocardial electrograms. Individual PWA varied from a decrease of 5.2 mV to an increase of 2 mV (-63% to 267%). In 33 patients with active fixation leads, the implant PWA was 1.96 +/- 0.99 mV versus 2.4 +/- 1.4 mV after surgery. In 11 patients with passive fixation leads, the implant PWA was 2.8 +/- 1.9 mV versus 1.9 +/- 0.8 mV after surgery. The PWA change, measured as the difference between the postsurgical and implant PWA was 0.43 +/- 0.8 mV in active versus -0.86 +/- 1.6 mV in the passive fixation lead groups (P less than 0.05). Considerable change in individual P wave amplitude can therefore occur very early after pacemaker implantation. The direction differs significantly between active (predominantly positive) and passive fixation groups (predominantly negative). These data suggest that an adequate margin of safety is important when initially programming atrial sensitivity, particularly when using passive fixation leads.

Aged↗

Autonomic responses in chest pain syndromes as compared to normal subjects.

The heart rate response to standing, cough, hand grip, and deep breathing were examined in normal subjects and coronary artery disease patients (greater than 70% diameter narrowing). The heart rate responses to these maneuvers were reduced in coronary patients and in anginal patients with normal coronary angiograms, as compared to normals. Detection (with the heart rate response to standing) was determined by using an RR interval cutoff of 140 ms for males and 120 ms for females discriminated between normals and CAD patients. In men sensitivity was 0.58, specificity 0.87 and CCR 0.75, and in women sensitivity was 0.67, specificity 0.79 and CCR 0.75. These values are similar to those reported for ST segment depression in similar populations. When separating normals from those with 2 and 3 vessel disease--sensitivity is 0.67, specificity 0.87, predictive value 0.71 and CCR 0.80. The response to cough, hand grip, and deep breathing showed similar trends but had less specificity than the response to standing. Thus, the heart rate response to most autonomic maneuvers is blunted in subjects with coronary disease and in those with pain syndromes sent for coronary angiography. These findings need testing in larger populations but autonomic maneuvers fail to discriminate patients with coronary disease from those with normal angiograms presenting with chest pain syndromes.

Adult↗

The role of prostaglandins E and F in acalculous gallbladder disease.

Prostaglandins have been postulated to be involved in the formation of gallstones and the pain and inflammation of calculous gallbladder disease. This report evaluated prostaglandin E and F levels in patients with acalculous gallbladder disease. Control gallbladders were obtained from patients undergoing cholecystectomy during insertion of hepatic artery catheters for regional, hepatic chemotherapy. Patients without gallstones and with long-standing post-prandial biliary colic with abnormal cholecystokinin administration underwent cholecystectomy for chronic acalculous cholecystitis. A third group of patients underwent cholecystectomy for acute acalculous cholecystitis. Gallbladder mucosa and muscle were separated, and prostaglandin E and F concentrations in mucosal and muscle or mucosa were identified in gallbladders from patients with chronic acalculous cholecystitis compared to gallbladders from patients without biliary tract symptoms. In gallbladders from patients with acute acalculous cholecystitis a seven-fold increase in PGE production by muscle tissue and mucosal cells was found. The more histologically inflamed gallbladders had higher mucosal and muscle prostaglandin E concentrations than were found in less inflamed gallbladders. Prostaglandin F levels were not significantly changed or were decreased, resulting in a significant increase in the ratio of PGE/PGF in acutely diseased gallbladders when compared to normal gallbladders. Prostaglandin E may be a manipulatable intermediary in the sequence of events that results in the development of acute acalculous cholecystitis.

Acute Disease↗

Genetic regulation of light damage to photoreceptors.

Albino mice of different inbred strains have been exposed to constant fluorescent lighting at an illuminance level of 115-130 ft-c for intervals of 1-6 weeks. Under these conditions the photoreceptors in retinas of albino BALB/cByJ mice rapidly degenerate, whereas the photoreceptors in retinas of albino C57BL/6J-c2J mice are remarkably more resistant to light damage. F1 heterozygotes produced from these two strains display an intermediate degree of light-induced degeneration. These findings demonstrate that phenotypically identical populations with different genetic constitutions can show markedly different sensitivities to light, and that genetic factors must now be included as a determinant for the severity of light damage.

Animals↗

Identification of phase-specific antigenic fractions of Coxiella burnetti by enzyme-linked immunosorbent assay.

Antigenic fractions of Coxiella burnetii phase variants were identified with an enzyme-linked immunosorbent assay (ELISA). Immune sera from guinea pigs immunized with Formalin-inactivated phase I or phase II whole cells were used to measure the antigenic activity of whole cells and various soluble and particulate preparations. Phase-specific antigens of C. burnetii whole cells and fractions were compared by dose-response curves at different (antigen and antibody) dilutions. Water-soluble extracts prepared by meta-periodate, ether, and phenol extraction of phase I whole cells yielded antigenic fractions which reacted with anti-phase I antibodies. The extraction of phase I whole cells with dimethyl sulfoxide, trichloracetic acid, and Formalin yielded antigenic fractions which detected antibodies in both anti-phase I and -phase II sera. Interestingly, the trichloracetic acid extract of phase I whole cells also contained a component which bound nonimmune immunoglobulin. The sera of animals immunized with whole cells of the phase II Australian QD strain reacted with lipopolysaccharides of the phase I and phase II Nine Mile strains. Therefore, variations in lipopolysaccharide structure among phase variants of C. burnetii were detected as cross-reactions with immune sera from an interspecific strain. Comparisons of immunofluorescence, microagglutination, and the complement fixation assays with the ELISA indicated greater sensitivity and specificity of the ELISA for the measurement of phase-specific antigens and antibodies.

Animals↗

Humoral immune response to Q fever: enzyme-linked immunosorbent assay antibody response to Coxiella burnetii in experimentally infected guinea pigs.

The response of guinea pigs experimentally infected with Coxiella burnetii organisms, the etiologic agents of Q fever, was obtained by the measurement of fever, circulating infectious C. burnetii cells, and anti-C. burnetii antibodies. The detection of antibodies by the enzyme-linked immunosorbent assay (ELISA) and traditional methods against phase I whole cells, phase II whole cells, and phase I lipopolysaccharide (LPS-I) (a virulence marker for phase I cells) antigens in the serum samples of infected animals revealed marked differences between intrastrain phase variants. Animals infected with the phase I Nine Mile strain produced a concomitant increase in temperature, circulating infectious C. burnetii cells, and antibodies against phase II cells, phase I cells, and LPS-I. At 15 weeks, a challenge of phase I-infected animals with viable phase I cells resulted in anamnestic antibody responses to phase I cells and LPS-I but not to phase II cells. Infection of animals with the phase II Nine Mile strain produced antibodies against only phase II cells. The challenge of phase II-infected animals at 15 weeks with viable phase II cells resulted in anamnestic antibody responses to phase I and phase II cells but not to LPS-I. Suppression of anti-phase II responses by the phase I challenge was apparent with only the ELISA, because the immunofluorescence, microagglutination, and complement fixation assays were insensitive to these changes. The sensitivity and specificity of the ELISA with whole-cell and the LPS-I antigens in the detection of phase-specific antibody revealed that avirulent phase II cells induced an immune response to phase I antigenic epitopes. Although the avirulent phase II cells were rapidly cleared by the host immune responses, they were sufficiently infective to induce antibody responses to both phase variants. Thus, in the occurrence of Q fever, any conventional serological technique that uses only phase II antigens may not provide a true incidence of naturally acquired infection with both phase I and II C. burnetii organisms.

Agglutination Tests↗