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Biomedical subjects

R K Johnson

Publications and source records attributed to R K Johnson.

At least 19 recordsLinked to original sources

In-person vs telephone-administered multiple-pass 24-hour recalls in women: validation with doubly labeled water.

OBJECTIVE: To determine the accuracy of energy intakes estimated with the multiple-pass 24-hour recall method in women by conducting in-person and telephone interviews. Doubly labeled water measurements of total energy expenditure were used for validation. SUBJECTS: Thirty-five weight-stable women (mean age = 30 years, range = 19 to 46 years) participated. DESIGN: Total energy expenditure was measured over a 14-day period using the doubly labeled water method. During this time, 4 multiple-pass 24-hour recalls were obtained from the women (2 in-person, 2 by telephone) who were provided 2-dimensional food models to estimate portion sizes. The Food Intake Analysis System was used to analyze recall data. STATISTICAL ANALYSES: Paired t tests were conducted to examine differences between energy intake estimated from the telephone and in-person interviews. Agreement between the energy intake estimates from the telephone recalls and the in-person recalls was assessed using the technique of Bland and Altman. Paired t tests were used to compare energy intake estimated from the telephone and in-person recalls to total energy expenditure. RESULTS: No significant difference in mean daily energy intake was found between the telephone (2,253 +/- 688 kcal) and in-person (2,173 +/- 656 kcal) interviews (P = .36). However, the mean energy intake from each interview method was significantly lower than total energy expenditure (2,644 +/- 503 kcal) (P = .006 and .001, respectively). APPLICATIONS/CONCLUSIONS: Underreporting of energy intake was widespread in the sample. Although the multiple-pass 24-hour recall method did not generate a group measure of energy intake that was accurate or unbiased, the telephone-administered multiple-pass 24-hour recall was just as effective in estimating energy intake as the recall administered in-person. Dietetics professionals should be aware of the pervasive and serious problem of under-reporting of self-reported food intakes.

Adult↗

Changing eating and physical activity patterns of US children.

The number of US children who are overweight has more than doubled over the last decade. This change has broadened the focus of dietary guidance for children to address nutrient overconsumption and physical activity patterns. Total fat consumption expressed as a percentage of energy intake has decreased among US children. However, this decrease is largely the result of increased total energy intake in the form of carbohydrates and not necessarily due to decreased fat consumption. The majority of children aged 5-17 years are not meeting recommendations for Ca intakes. Much of this deficit is attributed to changing beverage consumption patterns, characterized by declining milk intakes and substantial increases in soft-drink consumption. On average, US children are not eating the recommended amounts of fruits and vegetables. US adolescents become less active as they get older, and one-quarter of all US children watch > or = 4 h television each day, which is positively associated with increased BMI and skinfold thickness. There is an urgent need in the USA for effective prevention strategies aimed at helping children grow up with healthful eating and physical activity habits to achieve optimal health.

Adolescent↗

Bioactive compounds from Combretum erythrophyllum.

A methanol extract of Combretum erythrophyllum showed inhibitory bioactivities in a yeast-based microtiter assay for DNA-damaging agents. Bioassay-guided fractionation of this extract yielded two known bioactive compounds, combretastatin A-1 and (-)-combretastatin, and two new bioactive glucosides, combretastatin A-1 2'-beta-D-glucoside (1) and combretastatin B-1 2'-beta-D-glucoside (2). The structures of the new compounds were assigned by (1)H and (13)C NMR, DEPT, HMQC, and HMBC spectra.

Antineoplastic Agents, Phytogenic↗

Isolation and biochemical characterization of a new topoisomerase I inhibitor from Ocotea leucoxylon.

In a continuation of our search for potential tumor inhibitors from plants, we found that a crude extract from Ocotea leucoxylon showed selective activity typical of inhibitors of the enzyme topoisomerase I in a yeast assay for DNA-damaging agents. Using a bioassay-directed fractionation approach, the major bioactive compound was isolated and identified as the known aporphine alkaloid dicentrinone (4); the inactive alkaloid dicentrine (3) was also isolated. Compound 4 showed selective bioactivity against the rad52 repair-deficient yeast strain RS322 (IC(12) 49 microg/mL) and was inactive against the rad52- and topo1-deficient strain RS321 (IC(12) > 2000 microg/mL) and against the repair-proficient strain RJ03 (IC(12) > 2000 microg/mL). Biochemical studies with recombinant human topoisomerase I indicated that dicentrinone (4) is an inhibitor of the human enzyme. Colony formation studies suggest that it is weakly cytotoxic, but that its mechanism of toxicity differs from that of camptothecin and its derivatives.

Aporphines↗

Comparison of plasma FSH concentration in boars and gilts from lines selected for ovulation rate and embryonal survival, and litter size and estimation of (co)variance components for FSH and ovulation rate.

The objective of this research was to determine whether plasma concentration of FSH was genetically correlated with ovulation rate and thus was a useful trait for indirect selection. Blood samples were collected from 619 animals from five lines of pigs. Line I was selected for increased index of ovulation rate and embryonal survival, and Line C was its randomly selected control. Pigs sampled from Lines I and C were from generations 12 and 13. Pigs from three additional lines that were derived from eighth-generation pigs of Lines I and C also were used. These lines were Line C2, a randomly selected control derived from Line C, Line COL, derived from Line C, and Line IOL, derived from Line I; each of these lines was selected an additional five generations for increased ovulation rate and increased litter size. A single blood sample was collected from each pig between 46 to 63 (d 58), 86 to 98 (d 90), 110 to 133 (d 124), and 147 to 153 (d 150) d of age. The heritability of ovulation rate was .28 and heritabilities of plasma concentration of FSH at d 58, 90, 124, and 150 were .41, .25, .12, and 0, respectively. Genetic correlations between ovulation rate and d-58, d-90, and d-124 plasma concentration of FSH were .31, .23, and 0, respectively. Line I gilts had greater estimated breeding values for plasma concentration of FSH at d 58 and 90 than Line C gilts (P < .01). Line COL gilts had greater estimated breeding values for plasma concentration of FSH at d 58 than Line C2 gilts (P < .01). Line I boars had greater estimated breeding values for plasma concentration of FSH at d 90 than Line C boars (P < .05). Even though genetic correlations were low, selection for increased plasma concentration of FSH was estimated to be 93% as effective in changing ovulation rate as direct selection because selection for FSH can be practiced in both sexes. Thus, selection for increased plasma concentration of FSH seems to be a practical method for increasing ovulation rate in pig breeding programs without using laparoscopy.

Animals↗

Mammalian Chk2 is a downstream effector of the ATM-dependent DNA damage checkpoint pathway.

In response to DNA damage and replication blocks, cells activate pathways that arrest the cell cycle and induce the transcription of genes that facilitate repair. In mammals, ATM (ataxia telangiectasia mutated) kinase together with other checkpoint kinases are important components in this response. We have cloned the rat and human homologs of Saccharomyces cerevisiae Rad 53 and Schizosaccharomyces pombe Cds1, called checkpoint kinase 2 (chk2). Complementation studies suggest that Chk2 can partially replace the function of the defective checkpoint kinase in the Cds1 deficient yeast strain. Chk2 was phosphorylated and activated in response to DNA damage in an ATM dependent manner. Its activation in response to replication blocks by hydroxyurea (HU) treatment, however, was independent of ATM. Using mass spectrometry, we found that, similar to Chk1, Chk2 can phosphorylate serine 216 in Cdc25C, a site known to be involved in negative regulation of Cdc25C. These results suggest that Chk2 is a downstream effector of the ATM-dependent DNA damage checkpoint pathway. Activation of Chk2 might not only delay mitotic entry, but also increase the capacity of cultured cells to survive after treatment with gamma-radiation or with the topoisomerase-I inhibitor topotecan.

Alkylating Agents↗

Synthesis of furanonaphthoquinones with hydroxyamino side chains.

Several furanonaphthoquinones have shown useful activity in a yeast assay for DNA-damaging agents and cytotoxicity in mammalian cell culture assays. These results, together with the planar aromatic character of the furanonaphthoquinones, suggested that they might be acting as DNA intercalators. In an attempt to improve this activity, various analogues containing a hydroxyamino side chain have been synthesized. The analogues were prepared by standard methods, but some unexpected reactions were observed nonetheless. Thus, 8-formyl-5-methoxy-4,9-dihydronaphtho[2,3-b]furan-4,9-dione (24) showed an unusual reactivity toward reductive amination, with the reaction proceeding further to give one of two different cyclized products, depending on the amination reagent used. Bioassay results indicated that only simple furanonaphthoquines showed activity in a yeast assay for DNA-damaging agents; compounds with a substituted hydroxyamino side chain were uniformly inactive in this assay. Most of the compounds with a substituted hydroxyamino side chain on the furan ring did, however, show cytotoxicity, although none of them was any more active than the simple aldehyde 2-formyl-4, 9-dihydronaphtho[2,3-b]furan-4,9-dione (14). This evidence tends to suggest that the furanonaphthoquinones do not serve primarily as DNA intercalators, because if this were the case, they would have been expected to show an increased activity on conversion to their hydroxyamino side chain derivatives.

Animals↗

Responses in ovulation rate, embryonal survival, and litter traits in swine to 14 generations of selection to increase litter size.

Eleven generations of selection for increased index of ovulation rate and embryonal survival rate, followed by three generations of selection for litter size, were practiced. Laparotomy was used to count corpora lutea and fetuses at 50 d of gestation. High-indexing gilts, approximately 30%, were farrowed. Sons of dams in the upper 10% of the distribution were selected. Selection from Generations 12 to 14 was for increased number of fully formed pigs; replacements were from the largest 25% of the litters. A randomly selected control line was maintained. Responses at Generation 11 were approximately 7.4 ova and 3.8 fetuses at 50 d of gestation (P < .01) and 2.3 fully formed pigs (P < .01) and 1.1 live pigs at birth (P < .05). Responses at Generation 14 were three fully formed pigs (P < .01) and 1.4 live pigs (P < .05) per litter. Number of pigs weaned declined (P < .05) in the index line. Total litter weight weaned did not change significantly. Ovulation rate and number of fetuses had positive genetic correlations with number of stillborn pigs per litter. Significantly greater rate of inbreeding and increased litter size at 50 d of gestation in the select line may have contributed to greater fetal losses in late gestation, greater number of stillborn pigs, and lighter pigs at birth, leading to lower preweaning viability. Heritabilities of traits were between 8 and 25%. Genetic improvement programs should emphasize live-born pigs and perhaps weight of live-born pigs because of undesirable genetic relationships of ovulation rate and number of fetuses with numbers of stillborn and mummified pigs and because birth weight decreased as litter size increased.

Animal Husbandry↗

Diaminobenzidine as a myelin stain in semithin plastic sections.

A diaminobenzidine (DAB) stain for myelin in glutaraldehyde fixed, osmicated, semithin epoxy sections is described. One or 1.5 microm sections, dried onto slides, are first etched with a 1:2 dilution of saturated sodium ethoxide:absolute ethanol, then incubated in 0.05% aqueous DAB with 0.01% hydrogen peroxide. DAB specifically stains osmium fixed myelinated nerve fibers. This permits high resolution light microscopic study of myelinated nerve fibers in semithin sections of tissues that also can be studied by electron microscopy.

3,3'-Diaminobenzidine↗

Madman in the OR.

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General Surgery↗

Concentrations of gonadotropins, estradiol and progesterone in sows selected on an index of ovulation rate and embryo survival.

The objective of this study was to determine concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4) and 17beta-estradiol (E2) in sows from a line selected on an index which emphasized ovulation rate (Select) and from a control line. A further classification of the sows in each line was made according to the estimated number of ovulations during an estrous cycle. Sows in the Select line were ranked into a high (HI) or low group (LI) when their estimated number of ovulations were 25 or more and 14 to 15, respectively. Sows of the control line were classified into groups as high (HC) or low (LC) when the estimated values for ovulation rate were 14-15 and 8-9 ovulations, respectively. Blood samples were collected every 12 h during a complete estrous cycle and samples were analyzed for concentrations of FSH and LH. Samples collected every 24 h were assayed for P4 and E2. Mean concentrations of FSH, LH, P4 and E2 did not differ (P > 0.10) between lines or between HI and LI or HC and LC groups. Selection of pigs for ovulation rate and embryonal survival did not affect concentrations of FSH, LH, P4 and E2 in sows during the estrous cycle.

Animals↗

A small, nonpeptidyl mimic of granulocyte-colony-stimulating factor [see commetns].

A nonpeptidyl small molecule SB 247464, capable of activating granulocyte-colony-stimulating factor (G-CSF) signal transduction pathways, was identified in a high-throughput assay in cultured cells. Like G-CSF, SB 247464 induced tyrosine phosphorylation of multiple signaling proteins and stimulated primary murine bone marrow cells to form granulocytic colonies in vitro. It also elevated peripheral blood neutrophil counts in mice. The extracellular domain of the murine G-CSF receptor was required for the activity of SB 247464, suggesting that the compound acts by oligomerizing receptor chains. The results indicate that a small molecule can activate a receptor that normally binds a relatively large protein ligand.

Animals↗

Differential effects of camptothecin derivatives on topoisomerase I-mediated DNA structure modification.

The effects of eleven camptothecin derivatives on calf thymus topoisomerase I-mediated cleavage of synthetic DNA duplex have revealed that the A ring of camptothecin is very important for its biochemical activity. Depending on the type, number, and location of substituents, highly active or inactive analogues were obtained. The persistence of CPT-induced topoisomerase I-DNA covalent binary complexes was investigated by using as substrates DNA containing several good topoisomerase I cleavage sites, or else a synthetic DNA duplex of defined structure with a single high-efficiency cleavage site. The ligation kinetics at a given topoisomerase I cleavage site were sometimes quite different in the presence of CPT derivatives whose structures were closely related. Even in the presence of a single CPT analogue, topoisomerase I-DNA covalent binary complexes underwent ligation with different kinetics, presumably reflecting a dependence on DNA sequences flanking the individual topoisomerase I cleavage sites. Individual camptothecin derivatives also exhibited a spectrum of inhibitory potentials in blocking the topoisomerase I-mediated rearrangement of branched, nicked, and gapped DNA duplex substrates; in some cases the potencies of inhibition observed in these assays for individual camptothecin analogues were quite different than those determined for stabilization of the unmodified DNA-topoisomerase I binary complex.

Camptothecin↗