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Biomedical subjects

T Foley

Publications and source records attributed to T Foley.

At least 19 recordsLinked to original sources

Characterization of chloramphenicol and florfenicol resistance in Escherichia coli associated with bovine diarrhea.

Florfenicol, a veterinary fluorinated analog of thiamphenicol, is approved for treatment of bovine respiratory pathogens in the United States. However, florfenicol resistance has recently emerged among veterinary Escherichia coli isolates incriminated in bovine diarrhea. The flo gene, which confers resistance to florfenicol and chloramphenicol, has previously been identified in Photobacterium piscicida and Salmonella enterica serovar Typhimurium DT104. The flo gene product is closely related to the CmlA protein identified in Pseudomonas aeruginosa. The cmlA gene confers nonenzymatic chloramphenicol resistance via an efflux mechanism. Forty-eight E. coli isolates recovered from calves with diarrhea, including 41 that were both chloramphenicol and florfenicol resistant, were assayed for the presence of both flo and cmlA genes. Forty-two of the 44 isolates for which florfenicol MICs were > or =16 microg/ml were positive via PCR for the flo gene. All E. coli isolates for which florfenicol MICs were < or =8 microg/ml were negative for the flo gene (n = 4) Twelve E. coli isolates were positive for cmlA, and chloramphenicol MICs for all 12 were > or =32 microg/ml. Additionally, eight isolates were positive for both flo and cmlA, and both florfenicol and chloramphenicol MICs for these isolates were > or =64 microg/ml. DNA sequence analysis of the E. coli flo gene demonstrated 98% identity to the published GenBank sequences of both serovar Typhimurium flo(St) and P. piscicida pp-flo. The flo gene was identified on high-molecular-weight plasmids of approximately 225 kb among the majority of florfenicol-resistant E. coli isolates. However, not all of the florfenicol-resistant E. coli isolates tested contained the large flo-positive plasmids. This suggests that several of the E. coli isolates may possess a chromosomal flo gene. The E. coli flo gene specifies nonenzymatic cross-resistance to both florfenicol and chloramphenicol, and its presence among bovine E. coli isolates of diverse genetic backgrounds indicates a distribution much wider than previously thought.

Animals↗

External resorption presenting as an intracoronal radiolucent lesion in a pre-eruptive tooth.

A large intracoronal radiolucent lesion in an unerupted permanent molar was found during the routine assessment of a young male Caucasian prior to orthodontic treatment. The tooth was extracted. Histological examination indicated the lesion was caused by external resorption. The defect extended widely into the enamel and dentine, and was repaired in part by bone. The pulp chamber was not involved. The aetiology of these lesions is often obscure but in this case it appeared to have originated in the floor of two developmental pits on the occlusal surface of the tooth.

Child↗

Adult height in growth hormone (GH)-deficient children treated with biosynthetic GH. The Genentech Growth Study Group.

Near-adult height (AH) was determined in 121 children (72 males and 49 females) with GH deficiency (GHD) who were prepubertal when they began treatment with recombinant DNA-derived preparations of human GH. AH as a SD score was -0.7 +/- 1.2 (mean +/- SD), significantly greater than the pretreatment height SD score (-3.1 +/- 1.2), the predicted AH SD score (-2.2 +/- 1.2; Bayley-Pinneau method), and the height SD score at the start of puberty (-1.9 +/- 1.3). In contrast to studies of GH treatment outcome, which used pituitary-derived GH (pit-GH) in lower doses, we found that males did not have a higher AH SD score than females, spontaneous puberty did not diminish AH, and AH was significantly greater than that predicted at the start of GH treatment. In a multiple regression equation, the statistically significant variables (all P < 0.0001) related to AH (r2 = 0.70) were the following: duration of treatment with GH, sex (males were taller than females, as expected for the normal population), age (younger children had a greater AH) and height at the start of GH, and growth rate during first year of GH. For the AH SD score (r2 = 0.47), pretreatment predicted AH, duration of GH, and bone age delay were significant (P < 0.0002) explanatory variables. Bone age delay (chronological age-bone age) had a negative impact on the AH SD score. Target height, etiology of GHD, previous treatment with pituitary GH, and the presence or absence of spontaneous puberty did not significantly improve the prediction of AH. Early diagnosis of GHD and continuous treatment with larger doses of GH to near AH should improve the outcome in children with short stature due to GHD.

Body Height↗

Eye movement desensitization of public-speaking anxiety: a partial dismantling.

Forty college students suffering from public speaking anxiety and having experienced a specific traumatic speech-related event were exposed to either a standard EMD protocol with eye movements; a moving audio stimulus in place of the eye movements; a protocol with eyes resting on the hands in place of the eye movement, or a no-treatment control condition. The results revealed that EMD is comparable in limited effectiveness to the other procedures and that the eye movements are not a crucial component of the treatment with this population.

Adolescent↗

Characterization of a monoclonal antibody (HB-22) and development of an ELISA for human apolipoprotein A-I.

We describe the production and characterization of a high-affinity monoclonal antibody, HB-22, for apolipoprotein (apo) A-I, a major protein of human high-density lipoproteins (HDL). Including Tween 20 in the reaction mixture increased the binding capacity of HB-22 to apo A-I. HB-22 showed monospecific reactivity with HDL or apo A-I, displaying no cross-reactivity with apo A-II, intermediate-, low-, or very-low-density lipoproteins. Immunoaffinity columns with HB-22 (in the absence of Tween 20) showed an immunosorbent capacity of 80 micrograms of apo A-I per milligram of antibody. The immunosorbent capacity of HB-22 for apo A-I was similar in plasma samples from normolipidemic, hypercholesterolemic, or hypertriglyceridemic patients. Comparative binding studies demonstrated that compared with other available monoclonal apo A-I antibodies, HB-22 had the greatest apparent affinity for binding to HDL. A competitive ELISA developed by utilizing HB-22 could detect as little as 20 ng of apo A-I in the reaction mixture. The intra- and interassay CVs of the ELISA were 5.4% and 9.5%, respectively.

Animals↗

Gender-biased language in learning disability textbooks.

The purpose of our study was to explore whether gender-biased language exists in two editions of popular learning disabilities (LD) textbooks by a female author and a male author. First, we examined whether gender-biased language (as measured by the frequency of gender-specific pronouns) was present in LD textbooks as a function of the author's gender and discovered that the male author exhibited approximately a 7:1 ratio in his use of masculine to feminine pronouns. In contrast, the female author demonstrated no significant difference in her use of gender-specific pronouns. Second, we investigated whether the degree of gender-biased language has changed in different editions over the past 5 years and found a significant change only in the use of neutral pronouns by the male author. The existence of gender-biased special education textbooks raises the issue of the possible influence that such material may have on professional expectations concerning students with LD. In a field in which males are placed in LD programs in disproportionate numbers, the responsibilities of both authors and publishers are discussed, as their actions may have an impact on female students' rights in accessing appropriate educational services.

Adolescent↗

Thyroid hormone metabolism under extreme body exercises.

In two runs over a distance of 75 km and 45 km as well as in a marathon (42.2 km), the thyroid hormones (T4, T3 and TSH in all runs and rT3 in two of them) were determined before and after the runs. The performance of the runners was analyzed, with their age considered in the analysis. We found that the changes of thyroid hormones are characterized by individual differences. Younger runners and those with better performances had significantly higher T4 serum concentrations and unaltered T3 and TSH concentrations, whereas older sportsmen and those with worse performances reacted to the stressful stimulus with a significant decrease of T4, T3 and TSH. This response was interpreted as an exhaustion reaction, i.e. reduced adaptation responses of thyroid hormone metabolism. Therefore, these investigations provide an assessment of the state of fitness, the optimization of training and the avoidance of damage caused by overly stressful physical exercise

Adult↗

Interlaboratory testing of the transferability of a candidate reference method for total cholesterol in serum.

In 1975 the Centers for Disease Control, in cooperation with the American Association for Clinical Chemistry Cholesterol Reference Method Study Group, began an investigation to develop a reference method for total cholesterol. Five potential reference methods were compared with the definitive method developed by the National Bureau of Standards before the chemical method of Abell et al. (J Biol Chem 1952;195:357-66) was selected as the recommended reference method. Because acceptance of a proposed reference method depends so greatly on the method's capability for transfer to other laboratories by written specifications and instructions, a transferability testing study was designed and conducted with 14 laboratories. The study consisted of preliminary testing of readiness of equipment, reagents, and personnel followed by transferability testing with eight runs on 10 serum pools. Laboratoires that did not meet readiness specifications had higher CVs in the transferability testing. The study demonstrated that the proposed method permits laboratories to attain a CV of less than 1.5% for one laboratory and of less than 3.0% among laboratories. The mean percent bias value was less than 1.0% for six of the 14 laboratories, less than 1.5% for 12, and less than 3.0% for all 14 laboratories.

Calibration↗

Induction of delta-Aminolevulinic Acid Synthase Activity and Inhibition of Heme Synthesis in Euglena gracilis by N-Methyl Mesoporphyrin IX.

N-Methyl mesoporphyrin IX, an inhibitor of heme synthesis, increases extractable delta-aminolevulinic acid (ALA) synthase activity when administered to growing cultures of Euglena gracilis Klebs strain Z Pringsheim in micromolar concentrations. Wild-type light-grown green cells and white aplastidic cells exhibited 2.8-fold and 1.8-fold increases, respectively, in ALA synthase activity within five to six hours after incubation with 4 x 10(-6) molar N-methyl mesoporphyrin IX. Protoheme levels were decreased and (59)Fe incorporation into heme was inhibited by N-methyl mesoporphyrin IX, indicating that, as in animal cells, N-methyl mesoporphyrin IX acts specifically to block iron insertion into protoporphyrin IX. Chlorophyll synthesis in wild-type cells was not affected within the first 6 hours after administration of N-methyl mesoporphyrin IX.

Journal Article↗

delta-Aminolevulinic Acid Synthase of Euglena gracilis: Regulation of Activity.

delta-Aminolevulinic acid (ALA), a key precursor of the tetrapyrroles heme and chlorophyll, is capable of being synthesized by two different routes in cells of the unicellular green alga Euglena gracilis: from the intact carbon skeleton of glutamate, and via the condensation of glycine and succinyl CoA, mediated by the enzyme ALA synthase. The regulatory properties of ALA synthase were examined in order to establish its role in Euglena.Partially purified Euglena ALA synthase, unlike the case with the bacterial or animal-derived enzyme, does not exhibit allosteric inhibition by the tetrapyrrole pathway products heme, protoporphyrin IX, and porphobilinogen, at concentrations up to 100 micromolar.In aplastidic mutant cells, extractable ALA synthase activity is constant during exponential growth, and decreases to low levels as the cells reach the stationary state. Rapid exponential decline of ALA synthase (t(1/2) = 55 min) occurs after administration of 43 micromolar cycloheximide, but not 6.2 millimolar chloramphenicol. These results suggest that, as in other eukaryotic cells, ALA synthase is synthesized on cytoplasmic ribosomes and is subject to rapid turnover in vivo.Extractable ALA synthase activity increases 2.5-fold within 6 hours after administration of 100 millimolar ethanol, a stimulator of mitochondrial development, and 4.5-fold within 12 hours after administration of 1 millimolar 4,6-dioxoheptanoic acid, which blocks ALA utilization, suggesting that activity is controlled in vivo by a feedback induction-repression mechanism, coupled with rapid enzyme turnover.In heterotrophically grown wild-type cells, low levels of ALA synthase rapidly increase 4.5-fold within 12 hours after cells are transferred from the light to the dark, and decrease exponentially (t(1/2) = 75 min) when cells are transferred from the dark to light. The dark levels are equal to those in light- or dark-grown aplastidic mutant cells. The low level occurring in light-grown wild-type cells is not altered by the presence of 10 micromolar 3-(3,4-dichlorophenyl)-1,1-dimethylurea, which blocks photosynthetic O(2) production. The decrease that occurs on dark-to-light transfer can be diminished by 12- or 24-hour prior incubation with 6.2 millimolar chloramphenicol, which also retards chlorophyll synthesis after the transfer to light.The positive relationship of ALA synthase activity to degree of mitochondrial expression, and the inverse relationship to plastid development and chlorophyll synthesis, suggests that ALA synthase functions to provide precursors to nonplastid tetrapyrroles in Euglena. In light-grown, wild-type cells, the diminished levels of ALA synthase may be due to the ability of developing plastids to export heme or a heme precursor to other cellular regions, which thereby supplants the necessity for ALA formation via the ALA synthase route.

Journal Article↗

delta-Aminolevulinic Acid Formation from gamma,delta-Dioxovaleric Acid in Extracts of Euglena gracilis.

gamma,delta-Dioxovaleric acid (DOVA) has been proposed as a precursor to heme and chlorophyll in plants and algae. DOVA transaminase activity was found in extracts of the unicellular green alga Euglena gracilis Klebs strain Z Pringsheim. Optimum conversion of DOVA to delta-aminolevulinic acid (ALA) occurred at pH 6.8. ALA formation was linear with time for at least 30 minutes at 37 degrees C and was proportional to amount of cell extract in the incubation mixture. Boiled cell extract was inactive. DOVA transaminase from either wild-type or aplastidic derivative strain W(14)ZNaIL ran as a single band in agarose gel permeation chromatography, with a calculated molecular weight of 98,000 +/- 3,000. l-Glutamic acid was the most effective amino donor. d-Glutamic acid was inactive. K(m) values for l-glutamic acid and DOVA were 11 and 1.1 millimolar, respectively. Pyridoxal phosphate stimulated activity maximally at 30 micromolar, and (aminooxy)acetate was strongly inhibitory. Glyoxylic acid was a competitive inhibitor with respect to DOVA, with an inhibition constant of 0.62 millimolar. Wild-type and aplastidic cells vielded equal activity, 31 +/- 1 nanomoles ALA per 30 minutes per 10(7) cells, whether grown in light or dark. DOVA transaminase could not be separated from glyoxylate transaminase activity by agarose gel permeation or diethylaminoethyl-cellulose column chromatography. In all fractions, glyoxylate transaminase activity was at least 75 times greater than DOVA transaminase activity. DOVA transamination appears to be catalyzed by glyoxylate transaminase, and not to be of physiological significance with respect to chlorophyll synthesis in Euglena.

Journal Article↗

delta-Aminolevulinic acid synthase from Euglena gracilis.

delta-Aminolevulinic acid (ALA) synthase [succinyl-CoA:glycine C-succinyltransferase (decarboxylating), EC 2.3.1.37] activity was detected in cell extracts of the unicellular green flagellate alga Euglena gracilis. The enzyme was identified by substrate and cofactor requirements, and activity was proportional to number of cells extracted and duration of incubation. The incubation product was spectrophotometrically and chromatographically identical to ALA. ALA synthase activity is present in two wild-type strains, Z and bacillaris, and in nongreening, aplastidic strains derived from them. When grown in the dark, wild-type strains have amounts of ALA synthase activity equal to the amounts in their aplastidic derivative strains. Growth in the light or dark does not affect the level of ALA synthase activity in the aplastidic strains, but the wild-type strains have only one-third as much activity when grown in the light. We propose that ALA synthase is responsible for nonplastid tetrapyrrole biosynthesis in Euglena.

5-Aminolevulinate Synthetase↗

Heparan sulphate sulphotransferase. Properties of an enzyme from ox lung.

A heparan sulphate sulphotransferase was partially purified from an ox lung homogenate by (NH(4))(2)SO(4) precipitation. Various glycosaminoglycans were assayed as sulphate acceptors with this enzyme. The highest acceptor activity was obtained with desulphated heparin and heparan sulphate, which indicates that sulphate transfer may be to free amino groups of the substrate. Some heparan sulphate was (35)S-labelled by incubation with the enzyme and re-isolated. On treatment of this heparan [(35)S]sulphate with nitrous acid and separation of the degradation products on Sephadex G-15, a major peak of radioactivity was obtained, and identified as [(35)S]sulphate by high-voltage electrophoresis at pH5.3. The [(35)S]sulphate is believed to be derived from N-[(35)S]sulphated groups of heparan [(35)S]-sulphate. That the ox lung preparation contained an N-sulphotransferase was confirmed by the isolation of 2-deoxy-2-[(35)S]sulphoamino-d-glucose as the major product from the flavobacterial degradation of heparan [(35)S]sulphate.

Animals↗