Comparison of a computerized physical activity recall with a triaxial motion sensor in middle-school youth.

PURPOSE: To develop and test a computerized activity recall (CAR) for capturing activity and energy expenditure (EE) in youth and to further our understanding of the use of the three-dimensional accelerometer (Tritrac) for studying activity and EE. METHODS: Forty-five students (25 girls and 20 boys) in grade 6-8, completed 5 consecutive days of the CAR and were randomly assigned to wear the Tritrac during 1 day in which they also completed the CAR. Twenty-two subjects also repeated 5 d of the CAR and 1 d of Tritrac monitoring 1-2 wk later. RESULTS: The correlation between CAR and Tritrac for total EE was significant (r = 0.510, P = 0.0003). However, the total EE computed from the Tritrac was significantly greater than the CAR (1941 +/- 273 kcal vs 1576 +/- 343 kcal (8.14 +/- 1.14 vs 6.60 +/- 1.44 MJ); P < 0.001). The EE of activities was similar for the Tritrac and CAR, approximately 670 kcal (2.80 MJ), suggesting that the difference between the two methods was their estimates of resting EE. Comparison of the CAR and Tritrac for classifying active and inactive subjects indicated that both methods similarly classified 35 of the 45 subjects or 78% (P = 0.0038). Although significant, the Kappa statistic (kappa = 0.398) indicated a low-level of agreement between methods. The Tritrac indicated 50.4 +/- 29.2 min.d-1 of activities of > or = 3 METs, whereas the CAR indicated 76.7 +/- 71.9 min.d-1 (P = 0.02). The week 1 vs 2 test-retest correlation was 0.0485 (P = 0.022) for the Tritrac and 0.820 (P = 0.0001) for the CAR. CONCLUSIONS: It appears that both methods have acceptable reliability. However, the validity of each method to measure total and activity EE remains unclear, as the Tritrac appears to overestimate resting energy expenditure, whereas the CAR overestimates total minutes of activity.

Effects of electromagnetic fields on molecules and cells.

Evidence suggests that cell processes can be influenced by weak electromagnetic fields (EMFs). EMFs appear to represent a global interference or stress to which a cell can adapt without catastrophic consequences. There may be exceptions to this observation, however, such as the putative role of EMFs as promoters in the presence of a primary tumor initiator. The nature of the response suggests that the cell is viewing EMFs as it would another subtle environmental change. The age and state of the cell can profoundly affect the EMF bioresponse. There is no evidence that direct posttranscription effects occur as a result of EMF exposure. Although transcription alterations occur, no apparent disruption in routine physiological processes such as growth and division is immediately evident. What is usually observed is a transient perturbation followed by an adjustment by the normal homeostatic machinery of the cells. DNA does not appear to be significantly altered by EMF. If EMF exposure is associated with an increased risk of cancer, the paucity of genotoxic effects would support the suggestion that the fields act in tumor promotion rather than initiation. The site(s) and mechanisms of interaction remain to be elaborated. Although there are numerous studies and hypotheses that suggest the membrane represents the primary site of interaction, there are also several different studies showing that in vitro systems, including cell-free systems, are responsive to EMFs. The debate about potential hazards or therapeutic value of weak electromagnetic fields will continue until the mechanism of interaction has been clarified.

Magnetic fields after translation in Escherichia coli.

Quantitative two-dimensional gel electrophoresis of proteins in E. coli exposed for 60 min to weak, pulsed magnetic fields (1.5 mT peak) show that numerous proteins are both increased and decreased by a factor of 2 or more. An increase in the levels of two proteins, the a subunit of DNA-dependent RNA polymerase and NusA, was confirmed by Western blot analysis.

Altered protein synthesis in a cell-free system exposed to a sinusoidal magnetic field.

This report describes a new approach for examining weak extremely low frequency (ELF) electric and magnetic field interactions with living systems that exploits a cell-free transcription/translation system derived from Escherichia coli. Using two-dimensional polyacrylamide gel electrophoresis we previously had determined that the level of the alpha subunit of RNA polymerase in intact E. coli was elevated by exposure to weak ELF magnetic fields. In this paper, plasmids containing the alpha, or both the beta,beta' subunits of the RNA polymerase from E. coli were placed into a cell-free expression system. When this transcription/translation system was exposed to a 72-Hz sinusoidal magnetic field in the range 0.07 to 1.1 mT (rms) for periods of 5 min to 1 h, expression was enhanced. Weaker fields must be applied longer to produce an effect. For 10 min of field exposure, the threshold for an effect is 0.1 mT. These experiments demonstrate that an intact membrane is not an absolute requirement for transducing magnetic bio-effects.

Periodicity of amide proton exchange rates in a coiled-coil leucine zipper peptide.

The two-stranded coiled-coil motif, which includes leucine zippers, is a simple protein structure that is well suited for studies of helix-helix interactions. The interaction between helices in a coiled coil involves packing of "knobs" into "holes", as predicted by Crick in 1953 and confirmed recently by X-ray crystallography for the GCN4 leucine zipper [O'Shea, E.K., Klemm, J.D., Kim, P.S., & Alber, T. (1991) Science 254, 539]. A striking periodicity, extending over six helical turns, is observed in the rates of hydrogen-deuterium exchange for amide protons in a peptide corresponding to the leucine zipper of GCN4. Protons at the hydrophobic interface show the most protection from exchange. The NMR chemical shifts of amide protons in the helices also show a pronounced periodicity which predicts a short H-bond followed by a long H-bond every seven residues. This variation was anticipated in 1953 by Pauling and is sufficient to give rise to a local left-handed superhelical twist characteristic of coiled coils. The amide protons that lie at the base of the "hole" in the "knobs-into-holes" packing show slow amide proton exchange rates and are predicted to have short H-bond lengths. These results suggest that tertiary interactions can lead to highly localized, but substantial, differences in stability and dynamics within a secondary structure element and emphasize the dominant nature of packing interactions in determining protein structure.

An increase in the negative surface charge of U937 cells exposed to a pulsed magnetic field.

Pulsed magnetic fields have been used to enhance healing of bone fractures and purportedly of lesions in soft tissue. However, their mechanism of action is poorly understood. We report changes in the plasma membrane of a nonadherent mammalian cell line, U937, which was exposed to a 25-pps magnetic field for 48 hours. Aqueous polymer two-phase partition studies showed that magnetic-field-exposed cells exhibited an increased negative surface charge but membrane hydrophobicity was not significantly altered. The observed increase in membrane electronegativity of exposed cells did not reflect a significant change in growth rate.

Folding of a peptide corresponding to the alpha-helix in bovine pancreatic trypsin inhibitor.

A short peptide corresponding to the alpha-helical region of BPTI shows partial folding in aqueous solution (pH 7) as judged by circular dichroism (CD). Folding is temperature and denaturant sensitive, and the peptide is monomeric. The difference CD spectrum, obtained from spectra at two temperatures, indicates that the peptide folds as an alpha-helix. Difference CD spectroscopy provides a sensitive assay for helix formation in peptides exhibiting small amounts of structure. Helix stability in this peptide shows a marked pH dependence which is consistent with stabilizing charged side-chain interactions with the helix dipole and/or salt bridge formation.

GrB deletion of the chorion locus of the silkmoth Bombyx mori. Localization of the left breakpoint and isolation of the deletion junction.

In the silkmoth Bombyx mori, choriogenesis occurs through the developmentally controlled deposition of several related classes of chorion proteins onto the oocyte by surrounding follicular cells. In the GrB mutant strain, a distinctive family of proteins (Hc) normally expressed late in choriogenesis, as well as several proteins of middle development specificity, are missing due to the deletion of the corresponding genes from the chorion locus. In addition, a smaller set of proteins normally confined to mid-choriogenesis is found to be prolonged in expression in homozygote mutant but not heterozygote individuals. To elucidate the molecular organization of the chorion locus in the GrB genotype, we scanned a part of the wild-type locus represented by a chromosomal walk of 270,000 bases through library screening and genomic DNA hybridizations using a series of unique probes. A chromosomal clone, GrB4, whose sequences showed the expected characteristics of the deletion junction, was isolated from a partial EcoRI library of mutant genomic DNA. Through comparative hybridizations, mapping and sequencing, the precise location of one of the deletion breakpoints was identified on one of the clones mapping in the characterized part of the wild-type locus. Attempts to locate the other breakpoint in wild-type DNA and to extend the structural characterization past the deletion junction through chromosomal walking were unsuccessful, due to the apparent absence of these sequences from libraries of wild-type and mutant genomic DNA, respectively. Hybridizations of the deletion region on clone GrB4 to cDNA derived from follicular RNA indicate that no gene sequences are directly interrupted by the deletion, and reveal the presence of a gene sequence of unknown function 1000 to 5000 bases to the right of deletion junction.

Low frequency electric and magnetic fields have different effects on the cell surface.

There is a considerable controversy over the nature of weak electromagnetic-field effects in living organisms. Part of the controversy can be traced to a lack of understanding of whether electric or magnetic fields are involved in producing bioeffects. We find that both 60 Hz electric and magnetic fields alter the cell surface of Physarum polycephalum. Exposure to electric fields increases the negative charge on the cell surface while magnetic-field exposure decreases the hydrophobic character of the surface. These effects appear to be additive and independent of the waveform of the applied fields.

Differences in the surface properties of the mating types of Physarum polycephalum.

In the slime mold Physarum polycephalum, formation of a diploid plasmodium occurs when compatible haploid amoebae fuse. To study cell surface changes associated with the fusion process, a non-destructive method known as aqueous, two-phase partitioning was employed. Using a two-phase system of dextran and polyethylene glycol, we observed that the two mating types (RSD4 and MA185) have different partition coefficients and hence different surface properties. Based on their partitioning behavior, MA185 cells appear to have a more hydrophobic surface than RSD4 amoeba. The partition coefficient of both cell types decreased with time. If amoebae were maintained in culture until they encysted, differences in their surface were not detectable.

Pulsed magnetic fields alter the cell surface.

Pulsed magnetic fields (PMFS) are routinely used in the medical community to facilitate bone repair in clinical cases of non-union or pseudarthoses [(1984) Orth. Clin. No. Am. 15, 61-87]. Although this therapeutic regimen appears to be reasonably effective, the mechanism of action between specific PMFs and the target tissue remains unknown. Adding urgency to the need to understand the mechanism are a wide number of reports that have appeared which demonstrate that PMFs similar to those in clinical use can alter many basic physiological functions. We report that a 24 h exposure to PMFs alters the cell surface of Physarum polycephalum amoebae. Further, using the technique of aqueous two-phase partitioning, we present evidence for individual magnetic and electric field, cell surface effects.

Effects of sinusoidal 60-Hz electric and magnetic fields on ATP and oxygen levels in the slime mold, Physarum polycephalum.

We have previously reported that exposing the vegetative plasmodia stage of Physarum polycephalum to either individual or simultaneously applied electric and magnetic fields (45-75 Hz, 0.14-2.0 G, and 0.035-0.7 V/m) lengthens their mitotic cycle, depresses their rate of reversible shuttle streaming, and lowers their respiration rate. In this article we report the effects of simultaneously applied electromagnetic fields (60 Hz, 1.0 G, 1.0 V/m), electric fields only (60 Hz, 1.0 V/m), magnetic fields only (60 Hz, 1.0 G) on the haploid amoeba of Physarum exposed for 120-180 days. Statistically significant depressions (about 8-11%) in ATP levels were observed with all field conditions; however, respiration was significantly decreased only when amoebae were subjected to either combined fields or electric fields alone. Magnetic fields alone failed to induce a significant decrease in respiration.

Magnetic field effects on mitotic cycle length in Physarum.

Large plasmodia of Physarum polycephalum were formed from mixtures of micro-plasmodia grown in shaker cultures exposed to 2.0 G (rms), 75 Hz magnetic fields and non-exposed, control cultures. The exposed cultures had been grown continuously in the field and displayed a longer mitotic cycle than the controls. Mixed cultures display synchronous mitosis and a cycle length intermediate to the cycle lengths of exposed and control cultures. The cycle length of mixed cultures varied with the proportions of the mixture in a non-linear manner. The results are discussed in terms of several models.

A flow cytometry study of the cell cycle and of ploidy levels in Physarum polycephalum myxamoebae and plasmodia.

The cell cycle of Physarum polycephalum myxamoebae, as well as haploid and diploid and plasmodia, was analysed using laser microfluorometry. The data obtained indicate that the cell cycle of all strains examined are similar. The DNA profiles also indicate that a haploid ploidy level is still uncertain and show conclusively that spontaneous chromosome loss is a parameter of concern in research involving these organisms.

The nuclear acidic proteins from haploid and diploid cell states of Physarum polycephalum.

The free-living haploid gametes of the Myxomycete Physarum polycephalum are biflagellate phagocytic cells displaying gross morphological and biochemical differences as compared to the diploid plasmodium. The residual acidic chromatin and nucleolar proteins from gametes compatible for mating-type allele have been examined and compared to identical fractions from the diploid plasmodium. The proteins from both gamete mating types are identical yet major differences in the complement of these proteins occur following karyogamy and plasmodial formation. These differences occur in a fraction of acidic chromatin proteins previously shown to contain polypeptides which undergo dramatic changes during differentiation of the diploid plasmodium.