Leveling the playing field: Evaluation of a portable instrument for quantifying balance performance

Balance is a complex, sensorimotor task requiring an individual to maintain the center of gravity within the base of support. Quantifying balance in a reliable and valid manner is essential to evaluating disease progression, aging complications, and injuries in clinical and research settings. Typically, researchers use force plates to track motion of the center of gravity during a variety of tasks. However, limiting factors such as cost, portability, and availability have hindered postural stability evaluation in these settings. This study compared the “gold standard” for assessing postural stability (i.e., the laboratory-grade force plate) to a more affordable and portable assessment tool (i.e., BTrackS balance plate) in healthy young adults. Correlations and Bland-Altman plots between the center of pressure outcome measures derived from these two instruments were produced. Based on the results of this study, the measures attained from the portable balance plate objectively quantified postural stability with high validity on both rigid and compliant surfaces, demonstrated by thirty-five out of thirty-eight observed postural stability metrics in both surface conditions with a correlation of 0.98 or greater. The low cost, portable system performed similarly to the lab-grade force plate indicating the potential for practitioners and researchers to use the BTrackS balance plate as an alternative to the more expensive force plate option for assessing postural stability, whether in the lab setting or in the field.     

Explanation and prediction in vegetation science

A definition of vegetation science is given, spanning 6 levels of integration and stressing the interrelations among them. The problems of realism are discussed. The selection of levels is related to the adequate correspondence between conceptualization and research aims. Pattern and process are introduced as the central concepts of vegetation science. The perception of reality is dependent on the spatial and temporal scale chosen. The concept of noise is discussed in relation to stochasticity and randomness of events. Traces of essentialism are found both in classification of communities and habitat ecology. Classification is important, particularly the coexistence of alternative classification approaches. Organicism as a basis of vegetation research is rejected because the organismic view is inadequate on higher integration levels. The concept of function is redefined in a non-teleologic way.Present vegetation ecological research is inductivistic. One possible alternative to inductivism is falsificationism. The major domain of this approach is hypothesis testing, which will become more important. Progress can only be reached by a maximum degree of communication among scientific individuals.Predictive ecology is partly based on historic explanation, partly on complementary approaches. Characters of vegetation worthwhile to be predicted are listed and the necessary requirements for vegetation science to become predictive are discussed. A major requirement is the development of succession and life-history theory. A further elaboration of the individualistic concept will be a main task of vegetation science in the near future.     

A simple modification to improve the accuracy of methylation-sensitive restriction enzyme quantitative polymerase chain reaction

DNA digestion with endonucleases sensitive to CpG methylation such as HpaII followed by polymerase chain reaction (PCR) quantitation is commonly used in molecular studies as a simple and inexpensive solution for assessment of region-specific DNA methylation. We observed that the results of such analyses were highly overestimated if mock-digested samples were applied as the reference. We determined DNA methylation levels in several promoter regions in two setups implementing different references: mock-digested and treated with a restriction enzyme that has no recognition sites within examined amplicons. Fragmentation of reference templates allowed removing the overestimation effect, thereby improving measurement accuracy.     

Automatic determination of the handedness of single-particle maps of macromolecules solved by CryoEM

Single-Particle Analysis by Cryo-Electron Microscopy is a well-established technique to elucidate the three-dimensional (3D) structure of biological macromolecules. The orientation of the acquired projection images must be initially estimated without any reference to the final structure. In this step, algorithms may find a mirrored version of all the orientations resulting in a mirrored 3D map. It is as compatible with the acquired images as its unmirrored version from the image processing point of view, only that it is not biologically plausible.In this article, we introduce HaPi (Handedness Pipeline), the first method to automatically determine the hand of electron density maps of macromolecules solved by CryoEM. HaPi is built by training two 3D convolutional neural networks. The first determines α-helices in a map, and the second determines whether the α-helix is left-handed or right-handed. A consensus strategy defines the overall map hand. The pipeline is trained on simulated and experimental data. The handedness can be detected only for maps whose resolution is better than 5 Å. HaPi can identify the hand in 89% of new simulated maps correctly. Moreover, we evaluated all the maps deposited at the Electron Microscopy Data Bank and 11 structures uploaded with the incorrect hand were identified.     

An optimized and validated RP-HPLC/UV detection method for simultaneous determination of all-trans-Retinol (Vitamin A) and α-Tocopherol (Vitamin E) in human serum: Comparison of different particulate reversed-phase HPLC columns

A novel, simple and fast reversed-phase HPLC/UV method was developed, optimized for various chromatographic conditions, and validated according to international guidelines for simultaneous determination of all-trans-retinol and α-tocopherol in human serum using retinyl acetate as internal standard in the concentration of 0.5 μg/ml. A liquid-phase extraction was applied to the 250 μl of serum with n-hexane–dichloromethane mixture (70:30, v/v), in two steps, using ethanol–methanol mixture (95:5, v/v) for protein precipitation and BHT (butylated hydroxy toluene) as stabilizer for sample preparation. Both analytes were analyzed on Kromasil 100 C₁₈ column (150 mm × 4.6 mm, 5 μm), Brownlee analytical (Perkin Elmer) C₁₈ column (150 mm × 4.6 mm, 5 μm), and Supelco (Supelcosil) LC-18 column (150 mm × 3 mm, 3 μm), protected by a Perkin Elmer C₁₈ (30 mm × 4.6 mm, 10 μm; Norwalk, USA) pre-column guard cartridge, at 292 nm wavelength, using methanol–water (99:1, v/v), in isocratic mode as mobile phase applied at flow rate of 1.5 ml/min and 1 ml/min for both 5 μm and 3 μm columns, respectively. Complete separation of all the analytes was achieved in 3 and 6 min on 3 μm and 5 μm columns, respectively by injecting 20 μl of sample into the HPLC system by autosampler, keeping column oven temperature at 25 °C. Different particulate reversed-phase chromatographic columns were evaluated in order to select the best column in terms of sensitivity, selectivity, resolution and short run time of both the analytes and it was concluded that 3 μm columns are better to be used in clinical set up as well as in laboratories for the separation of these analytes in a shorter time as compared with 5 μm columns. The method was validated and applied for the analysis of all-trans-retinol and α-tocopherol in the serum of human volunteers.     

HPLC method with fluorescence detection for the quantitative determination of flaxseed lignans

We report a rapid and simple HPLC method with fluorescence detection for the quantification of the major flaxseed lignan, secoisolarisiresinol diglucoside (SDG) and its major metabolites. The method is specific for SDG, secoisolarisiresinol (SECO), enterodiol (ED) and entrolactone (EL) in rat serum. The assay procedure involves chromatographic separation using a Waters Symmetry C₁₈ reversed-phase column (4.6 mm × 150 mm, 5 μm) and mobile phase gradient conditions consisting of acetonitrile (0.1% formic acid) and water (0.1% formic acid). SDG extraction from serum requires the use of Centrifuge filters while SECO, ED and EL are extracted with diethyl ether. The organic layer is evaporated and reconstituted in 100 μL of mobile phase and 50 μL of reconstituted sample or filtrate is injected onto the column. Total run time is 25 min. Calibration curves are linear (r² ≥ 0.997) from 0.05 to 10 μg/mL for SDG and EL and 0.01–10 μg/mL for SECO and ED. Precision and accuracy are within USFDA specified limits. The stability of all lignans is established in auto-injector, bench-top, freeze–thaw and long-term stability at −80 °C for 30 days. The method's reasonable sensitivity and reliance on more widely available HPLC technology should allow for its straightforward application to pharmacokinetic evaluations of lignans in animal model systems such as the rat.     

Validation model for Raman based skin carotenoid detection

Raman spectroscopy holds promise as a rapid objective non-invasive optical method for the detection of carotenoid compounds in human tissue in vivo. Carotenoids are of interest due to their functions as antioxidants and/or optical absorbers of phototoxic light at deep blue and near UV wavelengths. In the macular region of the human retina, carotenoids may prevent or delay the onset of age-related tissue degeneration. In human skin, they may help prevent premature skin aging, and are possibly involved in the prevention of certain skin cancers. Furthermore, since carotenoids exist in high concentrations in a wide variety of fruits and vegetables, and are routinely taken up by the human body through the diet, skin carotenoid levels may serve as an objective biomarker for fruit and vegetable intake. Before the Raman method can be accepted as a widespread optical alternative for carotenoid measurements, direct validation studies are needed to compare it with the gold standard of high performance liquid chromatography. This is because the tissue Raman response is in general accompanied by a host of other optical processes which have to be taken into account. In skin, the most prominent is strongly diffusive, non-Raman scattering, leading to relatively shallow light penetration of the blue/green excitation light required for resonant Raman detection of carotenoids. Also, sizable light attenuation exists due to the combined absorption from collagen, porphyrin, hemoglobin, and melanin chromophores, and additional fluorescence is generated by collagen and porphyrins. In this study, we investigate for the first time the direct correlation of in vivo skin tissue carotenoid Raman measurements with subsequent chromatography derived carotenoid concentrations. As tissue site we use heel skin, in which the stratum corneum layer thickness exceeds the light penetration depth, which is free of optically confounding chromophores, which can be easily optically accessed for in vivo RRS measurement, and which can be easily removed for subsequent biochemical measurements. Excellent correlation (coefficient R = 0.95) is obtained for this tissue site which could serve as a model site for scaled up future validation studies of large populations. The obtained results provide proof that resonance Raman spectroscopy is a valid non-invasive objective methodology for the quantitative assessment of carotenoid antioxidants in human skin in vivo.     

Regression Calibration in Semiparametric Accelerated Failure Time Models

Summary In large cohort studies, it often happens that some covariates are expensive to measure and hence only measured on a validation set. On the other hand, relatively cheap but error‐prone measurements of the covariates are available for all subjects. Regression calibration (RC) estimation method ( Prentice, 1982 , Biometrika 69 , 331–342) is a popular method for analyzing such data and has been applied to the Cox model by Wang et al. (1997, Biometrics 53 , 131–145) under normal measurement error and rare disease assumptions. In this article, we consider the RC estimation method for the semiparametric accelerated failure time model with covariates subject to measurement error. Asymptotic properties of the proposed method are investigated under a two‐phase sampling scheme for validation data that are selected via stratified random sampling, resulting in neither independent nor identically distributed observations. We show that the estimates converge to some well‐defined parameters. In particular, unbiased estimation is feasible under additive normal measurement error models for normal covariates and under Berkson error models. The proposed method performs well in finite‐sample simulation studies. We also apply the proposed method to a depression mortality study.     

Validation of thermal composting process using olive mill solid waste for industrial scale cultivation of Agaricus bisporus

The production of a substrate containing destoned olive mill solid waste for the cultivation of Agaricus bisporus (Lange) Imbach on an industrial scale was studied. A standard mushroom compost (C) mainly made from straw and poultry manure was compared with the experimental compost (EC) containing the same ingredients as (C) but with added olive mill solid waste (10.6% w/w). Microbial indicators such as counts of heterotrophic microbes and actinomycetes were higher in EC than in C. In addition, compost selectivity as indicated by higher mushroom yield and biological efficiency of EC was higher than that of C. Market quality of the mushrooms produced in both C and EC were comparable. These findings support our work that olive mill solid waste can be used safely in thermal composting process to produce a selective substrate for industrial-scale cultivation of A. bisporus. This study also demonstrates an environmentally sustainable system to manage solid waste from olive oil extraction processes thus overcoming environmental pollution brought about by irrational disposal of the waste on farm lands.