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Biomedical subjects

T L Scott

Publications and source records attributed to T L Scott.

At least 19 recordsLinked to original sources

Microbial degradation of illicit drugs, their precursors, and manufacturing by-products: implications for clandestine drug laboratory investigation and environmental assessment.

Chemicals associated with clandestine drug laboratories are often disposed of covertly into soil, sewerage systems, or public waste management facilities. There are two significant issues relating to such dumps of materials; they might contain valuable evidence as to drug manufacture, and they might be a source of pollution. This study presents initial findings in relation to the impact microorganisms from environmental sources have upon drugs, their precursors, and manufacturing by-products. The aim of this study was to identify which chemicals associated with clandestine drug laboratories persist in the environment in order to allow forensic drug chemists to link discarded residues with the method of manufacture, and to allow the environmental impact of clandestine drug laboratories to be assessed accurately. When exposed to soil microorganisms, phenyl-2-propanone (P2P) was rapidly metabolized into mixtures of 1-phenyl-2-propanol, 1-phenyl-1,2-propanedione, 1-hydroxy-1-phenyl-2-propanone, 2-hydroxy-1-phenyl-1-propanone, and the two diastereoisomers of 1-phenyl-1,2-propanediol. On the other hand, when exposed under the same conditions, methylamphetamine sulphate (MAS) remained virtually unchanged. Implications relating to evidence gathering for forensic purposes and to environmental assessment of clandestine drug laboratories are discussed.

2-Propanol↗

Corn processing method in finishing diets containing wet corn gluten feed.

Two trials were conducted to determine the effect of corn processing method on performance and carcass traits in steers fed finishing diets containing wet corn gluten feed (WCGF). In Trial 1, 480 steer calves (303 kg initial BW) were fed eight finishing diets: 1) dry-rolled corn (DRC) without; and 2) with 32% (DM basis) WCGF; 3) steam-flaked corn (SFC) without; and 4) with WCGF; 5) a combination of DRC and SFC without WCGF; 6) finely-ground corn (FGC) with WCGF; 7) high-moisture corn (HMC) with WCGF; and 8) whole corn (WC) with WCGF. Feeding WC + WCGF increased (P < 0.10) DMI and decreased gain:feed compared with all other treatments. Feeding DRC + WCGF increased (P < 0.10) DMI and decreased (P < 0.10) gain:feed compared with treatments other than WC + WCGF. Steers on treatments that included WCGF gained similarly, regardless of corn processing method, and at a rate 6% faster (P < 0.10) than steers fed diets that did not include WCGF. Gain:feed did not differ among steers fed SFC, SFC + WCGF, SFC + DRC, and HMC + WCGF. Steers fed SFC or SFC + WCGF were more efficient (P < 0.10) than steers fed DRC or FGC + WCGF. In Trial 2, 288 steer calves (382 kg initial BW) were fed six finishing diets: 1) DRC without; and 2) with 22% (DM basis) WCGF; 3) SFC without; and 4) with WCGF; 5) finely rolled corn (FRC) with WCGF; and 6) HMC corn with WCGF. Steers fed DRC + WCGF or FRC + WCGF consumed more DM (P < 0.10) than steers fed DRC, SFC, or SFC + WCGF. Feed intake did not differ between steers fed SFC + WCGF and HMC + WCGF. All treatment groups receiving WCGF consumed more DM (P < 0.10) feed than steers fed DRC or SFC without WCGF. Steers fed SFC + WCGF gained 8% faster (P < 0.10), and steers fed DRC 9.5% slower (P < 0.10) than steers receiving all other treatments. Daily gains did not differ among other treatment groups. Steers fed SFC or SFC + WCGF gained 10% more (P < 0.10) efficiently than all other treatment groups. Feed efficiency did not differ among steers fed DRC, DRC + WCGF, FRC + WCGF, and HMC + WCGF. Estimates for the NEg of WCGF calculated from animal performance indicated that WCGF contained approximately 25.3% more energy when fed with SFC than when fed with DRC. In general, more intensively processing corn improved gain:feed in finishing diets containing WCGF.

Animal Feed↗

Effect of corn processing on starch digestion and bacterial crude protein flow in finishing cattle.

Six ruminally and duodenally cannulated yearling steers (523 kg) were used in a replicated 3 x 3 Latin square design experiment to study the effects of corn processing on nutrient digestion, bacterial CP production, and ruminal fermentation. Dietary treatments consisted of 90% concentrate diets that were based on dry-rolled (DRC), high-moisture (HMC), or steam-flaked (SFC) corn. Each diet contained 2.0% urea (DM basis) as the sole source of supplemental nitrogen. Each period lasted 17 d, with d 1 through 14 for diet adaptation and d 15 through 17 for fecal, duodenal, and ruminal sampling. Dry matter and OM intakes were similar for DRC and SFC but were approximately 15% higher (P < 0.05) for HMC. True ruminal OM digestibilities were 18 and 10% greater (P < 0.05) for HMC than for DRC or SFC, respectively. Ruminal starch digestibilities were similar between HMC and SFC and were approximately 19% greater (P < 0.05) than DRC. Postruminal OM digestibility was similar among treatments; however, postruminal starch digestibility was 15% greater (P < 0.05) for SFC than for DRC or HMC, which were similar. Total-tract DM and OM digestibilities were similar between HMC and SFC and were 4% greater (P < 0.05) than DRC. Likewise, total-tract starch digestibilities were similar between HMC and SFC and were 3% greater (P < 0.05) than DRC. Bacterial CP flow to the duodenum was 29% greater (P < 0.05) for HMC than for DRC or SFC, which were similar. Bacterial N efficiencies were similar among treatments. Based on bacterial CP flow from the rumen, we estimate that dietary DIP requirements are approximately 12% higher for HMC-based diets than for DRC or SFC-based diets, which were similar.

Animals↗

Photoinduced DNA cleavage by cyclopentadienyl metal complexes conjugated to DNA recognition elements.

[structure: see text]. DNA recognition elements have been attached to CpW(CO)3CH3 and CpW(CO)3Ph, which produce methyl and phenyl radicals that cleave DNA upon photolysis. The inclusion of binding moieties in 3 increases the efficiency but not the selectivity of strand scission over that seen in the simple unfunctionalized complex, while 11 cleaves preferentially at T sites within AT-rich tracts.

Cyclopentanes↗

Induction of calcitonin and calcitonin receptor expression in rat mammary tissue during pregnancy.

Human breast milk samples at early time points after parturition contain high levels of calcitonin (CT) in both normal and thyroidectomized mothers, suggesting that mammary tissue produces CT. Using blot hybridization and reverse-transcriptase polymerase chain (RT-PCR) analysis of rat mammary RNA we found that CT messenger RNA is induced at midpregnancy (day 12), remains elevated through late pregnancy (day 19) but then decreases before the day of birth. RIA of mammary CT revealed that levels increase from 0.3 ng/g tissue in nonpregnant animals to peak at 1.6 ng/g on day 19 and then decline after that, paralleling messenger RNA expression. Dilution profiles for extracted mammary CT showed close parallelism with monomeric rat CT. Plasma samples from thyroparathyroidectomized rats contained 10-20 pg/ml CT that did not increase during pregnancy, suggesting that mammary CT is not released into plasma but functions locally. Consistent with this, RT-PCR detected that the CT receptor C1a isoform is expressed in rat mammary tissues during both pregnancy and lactation. This is the first report that mammary tissue expresses both CT and the CT receptor during pregnancy, suggesting that CT may have a paracrine regulatory role in the mammary gland.

Animals↗

Health literacy among Medicare enrollees in a managed care organization.

CONTEXT: Elderly patients may have limited ability to read and comprehend medical information pertinent to their health. OBJECTIVE: To determine the prevalence of low functional health literacy among community-dwelling Medicare enrollees in a national managed care organization. DESIGN: Cross-sectional survey. SETTING: Four Prudential HealthCare plans (Cleveland, Ohio; Houston, Tex; south Florida; Tampa, Fla). PARTICIPANTS: A total of 3260 new Medicare enrollees aged 65 years or older were interviewed in person between June and December 1997 (853 in Cleveland, 498 in Houston, 975 in south Florida, 934 in Tampa); 2956 spoke English and 304 spoke Spanish as their native language. MAIN OUTCOME MEASURE; Functional health literacy as measured by the Short Test of Functional Health Literacy in Adults. RESULTS: Overall, 33.9% of English-speaking and 53.9% of Spanish-speaking respondents had inadequate or marginal health literacy. The prevalence of inadequate or marginal functional health literacy among English speakers ranged from 26.8% to 44.0%. In multivariate analysis, study location, race/language, age, years of school completed, occupation, and cognitive impairment were significantly associated with inadequate or marginal literacy. Reading ability declined dramatically with age, even after adjusting for years of school completed and cognitive impairment. The adjusted odds ratio for having inadequate or marginal health literacy was 8.62 (95% confidence interval, 5.55-13.38) for enrollees aged 85 years or older compared with individuals aged 65 to 69 years. CONCLUSIONS: Elderly managed care enrollees may not have the literacy skills necessary to function adequately in the health care environment. Low health literacy may impair elderly patients' understanding of health messages and limit their ability to care for their medical problems.

Aged↗

Dissemination of an evidence-based intervention manual to improve managed care mammography rates.

After collaborating with the Centers for Disease Control and Prevention to develop an evidence-based mammography intervention manual, the dissemination of the manual in a managed care setting was assessed. Using qualitative methods (in-depth interviews), factors were examined that motivated use of the manual by the quality improvement units of eight Prudential HealthCare health plans. Seven of the plans implemented more intensive interventions after having received the manual than in the previous year. Factors that facilitated use of the manual and implementation of recommendations included: internal motivation, support of senior level management, adequate resources (time, personnel, and funds) to support interventions, and the organization and content of the manual itself.

Aged↗

Organometallic photonucleases: a novel class of DNA-cleaving agents.

The first demonstration of DNA cleavage by an organic radical generated via homolysis of a metal-alkyl bond in a Cp-metal complex is presented. Irradiation of CpW(CO)3CH3 (1.5 molecules/base pair) produced methyl radical, giving single-strand cleavage of pBR322 DNA. This process was inhibited by the general radical trap cysteine and by TEMPO, which traps carbon radicals but not oxygen-centered radicals.

Cyclic N-Oxides↗

Uncoupling of Ca2+ transport from ATP hydrolysis activity of sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase.

In reconstituted rabbit skeletal muscle (Ca2+ + Mg2+)-ATPase proteoliposomes, Ca(2+)-uptake is decreased by more than 90% with T2 cleavage (Arg-198). However, no difference in the ATP dependence of hydrolysis activity is seen between SR and trypsin-treated SR. A large decrease in E-P formation and hydrolysis activity of the enzyme appear only at T3 cleavage, which represents the cleavage of A1 fragment to A1a + A1b forms. The disappearance of hydrolysis activity due to digestion is prior to the disappearance of E-P formation. No significant difference is found in the passive Ca2+ efflux between control SR and tryptically digested SR in the absence of Mg2+ + ruthenium red or in the presence of ATP. However, the passive Ca2+ efflux rate for tryptically digested SR is much larger than control SR in the presence of Mg2+ + ruthenium red. These results show that the Ca2+ channel cannot be closed after trypsin digestion of SR membranes by the presence of the Ca2+ channel inhibitors, Mg2+ and ruthenium red. In the reconstituted proteoliposomes, the Ca2+ efflux rates are the same regardless of digestion (T2); also, efflux is not affected by the presence or absence of Mg2+ + ruthenium red. These results indicate that T2 cleavage causes 'uncoupling' of the 'Ca(2+)-pump' from ATP hydrolytic activity. A theoretical model is developed in order to fit the extent of tryptic digestion of the A fragment of the (Ca2+ + Mg2+)-ATPase polypeptide with the loss of Ca(2+)-transport. Fits of the theoretical equations to the data are consistent with that Ca(2+)-transport system appears to require a dimer of the polypeptide (Ca2+ + Mg2+)-ATPase.

Adenosine Triphosphate↗

Distances between the functional sites of the (Ca2+ + Mg2+)-ATPase of sarcoplasmic reticulum.

Luminescence energy transfer measurements have been used to determine the distances between the two high affinity Ca2+ binding-transport sites of the (Ca2+ + Mg2+)-ATPase of skeletal muscle sarcoplasmic reticulum. The lanthanide Tb3+ situated at one high affinity Ca2+ site was used as the transfer donor, and acceptors at the other Ca2+ site were the lanthanides Nd3+, Pr3+, Ho3+, or Er3+. Terbium bound to the enzyme was excited directly with a pulsed dye laser. Analysis of the changes in the terbium luminescence lifetime due to the presence of the acceptor indicates that the distance between the Ca2+ sites is 10.7 A. The distance between the Ca2+ sites and the nucleotide-binding catalytic site was determined using Tb3+ at the Ca2+ sites and either trinitrophenyl nucleotides (TNP-N) or fluorescein 5-isothiocyanate (FITC) in the catalytic site as energy acceptors. The R0 values for the Tb-acceptor pairs are approximately 30 and approximately 40 A for TNP-N and FITC, respectively. The distance between Tb3+ at the Ca2+ sites and TNP-ATP at the nucleotide site is approximately 35 A and that between the Ca2+ sites and the FITC labeling site is approximately 47 A. Considerations of the molecular dimensions of the ATPase polypeptide indicate that while the two Ca2+ sites are close to each other, the Ca2+ sites and the nucleotide site are quite remote in the three-dimensional structure of the enzyme.

Animals↗

Distinction of the roles of the two high-affinity calcium sites in the functional activities of the Ca2+-ATPase of sarcoplasmic reticulum.

The effects of trypsin digestion and low temperature on Ca2+ binding and on Ca2+ activation of ATP hydrolysis by the high-affinity transport sites of the Ca2+, Mg2+-ATPase of sarcoplasmic reticulum were examined. Sarcoplasmic reticulum vesicles contain 0.7-1.1 high-affinity Ca2+ sites per 10(5) g sarcoplasmic reticulum with K = 3-5 X 10(5) M-1, as well as sites of lower affinity. The first cleavage of the ATPase with trypsin (TD1) has no effect on the binding properties of the high affinity sites. The second tryptic cleavage (TD2) decreases the affinity of the high sites to K = 3 X 10(4) M-1 with conservation of the total number of sites. The purified ATPase contains 1.6-2.0 high affinity Ca2+ sites per 10(5) g protein when measured at 23 degrees C, while at 0-4 degrees C there is approximately equal to 1 high-affinity (K = 5 - 10 X 10(5) M-1) affinity site and approximately equal to 1 intermediate-affinity (K = 3 X 10(4) M-1) site per 10(5) g. Trypsin digestion to the point of TD1 has no effect on either the number or the binding constants of the high-affinity sites. Upon TD2 cleavage, one of the sites is converted to the intermediate-affinity state, while the other remains at high affinity. After TD2 modification of the enzyme both of the sites are in the intermediate affinity state at 4 degrees C. On the basis of the binding data, several models for the roles of the Ca2+ sites in the activation of ATP hydrolysis are derived. The results are summarized by a scheme in which the two high-affinity Ca2+ sites are heterogeneous with respect to sensitivity to temperature and to TD2 modification. The results of this and a previous study [Scott, T. L. and Shamoo, A. E. (1982) J. Membr. Biol. 64, 137-144] indicate that while occupation of either of the two Ca2+ sites can stimulate ATP hydrolysis, the site which is sensitive to TD2 is essential for the coupling of hydrolysis to Ca2+ transport.

Adenosine Triphosphate↗

Luminescence studies of Tb3+ bound to the high affinity sites of the Ca2+-ATPase of sarcoplasmic reticulum.

Direct excitation of lanthanide luminescence with a pulsed dye laser has been used to probe the molecular environment of the high affinity sites of the sarcoplasmic reticulum Ca2+-ATPase. The direct excitation spectrum of Tb3+ bound to these sites has been determined and a luminescence lifetime of approximately 1 ms measured. Measurements of the difference in lifetime of the Tb X ATPase complex in H2O and D2O indicate that there are approximately 2 H2O molecules in the first coordination sphere of Tb3+ bound at the high affinity sites of the ATPase. The results are compared with the properties of Tb3+ binding to high affinity sites of other Ca2+ binding proteins. The binding constant of Tb3+ to the ATPase is in the range of 0.3-5.0 X 10(8) M-1 as inferred from the KI for inhibition of ATP hydrolysis, in agreement with a previous report (Highsmith, S. R., and Head, M. R. (1983) J. Biol. Chem. 258, 6858-6862). The values of the Ca2+ binding constant (approximately 2 X 10(6) M-1) and the cooperative nature (n = 1.9) of Ca2+ protection of Tb3+ inhibition indicate that Tb3+ and Ca2+ compete for the high affinity sites of the ATPase. The results demonstrate that directly-excited Tb3+ luminescence provides unique information on the environment of the Ca2+ binding-transport sites of the SR ATPase.

Animals↗

A fluorescence probe study of the phosphorylation reaction of the calcium ATPase of sarcoplasmic reticulum.

The fluorescent thiol reagent N-(1-anilinonaphthyl-4)maleimide (ANM) reacts covalently with the Ca2+ ATPase moiety of fragmented sarcoplasmic reticulum in two phases as determined by the increase of fluorescence intensity and optical density at 350 nm. In the rapid phase, 5.5 nmol of ANM reacts with 1 mg of fragmented sarcoplasmic reticulum protein. Assuming that 55% of the total membrane protein is the Ca2+ ATPase, this is equivalent to 1 mol of SH/10(5) g of ATPase, designated as SH1-ANM. ANM reacts with the second SH (SH2-ANM) at a much slower rate. Reaction of ANM with both SH1-ANM and SH2-ANM produces no inhibition of phosphoenzyme (EP) formation. Upon addition of Mg . ATP in the micromolar range, at [Ca2+] = 1 microM there is an increase in the fluorescence intensity of ANM attached to SH2-ANM, while the ANM attached to SH1-ANM does not respond to Mg . ATP. Under conditions in which there is no EP formation, there is no fluorescence change. Furthermore, the enhancement of ANM fluorescence produced by Mg . ATP is reversed by ADP as it reacts with EP to form ATP. Thus, it appears that the Mg . ATP-induced fluorescence increase reflects changes of enzyme conformation produced by EP formation.

Adenosine Diphosphate↗

Active calcium treatment transport via coupling between the enzymatic and the ionophoric sites of Ca2+ + Mg2+-ATPase.

The 20K dalton fragment of Ca2+ + Mg2+-ATPase obtained from th tryptically digested sarcoplasmic reticulum has been further purified using Bio-Gel P-100. This removed low-molecular-weight UV-absorbing and positive Lowry-reacting contaminants. The ionophoric activity of the 20K fragment in both oxidized cholesterol and phosphatidylcholine:cholesterol membranes is unaltered by this further purification. The 20K selectivity sequence in phosphatidylcholine:cholesterol membrane is Ba2+ greater than Ca2+ greater than Sr2+ greater than Mn2+ Mg2+. Digestion of intact sarcoplasmic reticulum vesicles with trypsin, which results in the dissection of the hydrolytic site (30K) from the ionophoric site (20K), is shown to disrupt energy transduction between ATP hydrolysis and calcium transport. This further implicates the 20K dalton fragment as a calcium transport site. These data and previous evidence are discussed in terms of a proposed model for the ATPase molecular structure and the mechanisms of cation transport in sarcoplasmic reticulum.

Adenosine Triphosphatases↗