Cross-Validation of Fat-Free Mass Estimated From Body Density Against Bioelectrical Resistance: Effects of Obesity and Gender

GORAN, MICHAEL I AND M ABU KHALED. Cross-validation of fat-free mass estimated from body density against bioelectrical resistance: effects of obesity and gender. Obes Res. The major purpose of this study was to examine whether estimates of body composition from bioelectrical resistance were systematically biased by obesity and/or gender (using hydrodensitometry as a comparison method). We compared fat-free mass (FFM) by bioelectrical resistance (BR) using 5 equations (Lukaski, Kushner, Rising, Khaled, and Segal) to FFM by hydrodensitometry (HD) in 20 lean men, 30 lean women, 33 obese men and 22 obese women. None of the BR equations was successfully cross-validated against FFM by HD in all 4 sub-groups. The Lukaski equation significantly underestimated FFM in all 4 groups by 2.7 to 4.7 kg; the Kushner equation significantly underestimated FFM by 2.0 to 2.9 kg except in obese women; the Rising equation significantly overestimated FFM in obese women (5.3 kg) and men (2.9 kg); the Khaled equation successfully predicted FFM in all groups except obese men; and the Segal equation successfully predicted FFM in all groups except lean men. In some groups, a portion of the discrepancy could be explained by bias originating from body fat. Analysis of our data by forward regression analysis demonstrated that height²/resistance, body weight, gender and suprailiac skinfold thickness provide the most accurate estimates of FFM (R²=0.92; SEE = 3.58kg) that are free of bias originating from gender and body fat. We conclude that the estimation of fat-free mass by BR is significantly influenced by gender and obesity. An alternative equation is proposed for estimating fat-free mass based on measurement of height²/resistance, body weight, gender and suprailiac skinfold thickness.     

An improved method for mass rearing Paratheresia claripalpis for biological control of lepidopteran stalk borers

An improved method for mass rearing Paratheresia claripalpis Van der Wulp (Diptera: Tachinidae), for biological control of lepidopteran stalk borers is described and compared to the commonly used method. The improved method was found to be superior by being less labour intensive and by producing 2.25 times more parasites from the same number of hosts.

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Amélioration de la technique d'élevage de masse de Paratheresia claripalpis pour la lutte biologique contre Diatraea saccharalis

Résumé Une technique d'élevage de massa de la tachinaire, P. claripalpis Van der Wulp, est décrite et comparée à la technique classique. Cette technique correspond à une amélioration puisqu'elle demande moins de travail et produit 2.25 fois plus de parasites pour le même nombre d'hôtes.

     

Sensitivity of bioelectrical impedance to detect changes in human body composition

The purpose of this study was to compare the estimates of lean body mass (LBM) and percent body fat (%BF), as predicted by bioelectrical impedance (BIA) and sum of skinfolds (SF), with those derived by hydrostatic weighing (HW) obtained before and after a 10-wk diet and exercise regimen. The experimental (E) group consisted of 17 healthy male subjects; 20 healthy males served as the control (C) group. Post hoc Scheffé contrasts computed on E group data indicated that, for both LBM and %BF, the Lukaski and Segal BIA equations, as well as the Durnin SF equation, derived mean values that were not significantly different (0.05 significance level) from HW in both pre- and postregimen conditions. For LBM, the same equations derived the following significant (P less than 0.01) correlation coefficients for both pre- and postregimen data: Lukaski, 0.87 and 0.85; Segal, 0.89 and 0.87; and Durnin, 0.90 and 0.88. For %BF, the correlation coefficients were slightly lower but remained statistically significant (P less than 0.01). The findings of this study suggest that the BIA method, by use of either the Lukaski or Segal prediction equations, is a valid means of predicting changes in human body composition as measured by the Siri transformation of body density.     

Colonization of Lutzomyia shannoni (Diptera: Psychodidae) utilizing an artificial blood feeding technique

Laboratory colonization of hematophagous insects must include an efficient method of blood feeding, preferably by artificial means. Strict rules for obtaining animal use permits, extensive animal maintenance costs, and indirect anesthesia effects on animal health warrant the development of an artificial membrane feeding technique for sand fly colonization in laboratories. An attempt was made to colonize Lutzomyia shannoni using an artificial blood feeding membrane to replace the use of live animals commonly used for sand fly blood‐feeding purposes. Lutzomyia shannoni readily fed through a pig intestine membrane exposed at an angle of 45°. However, it did not feed through a chicken skin membrane. Olfactory attractants were unable to improve blood‐feeding efficiency. Plaster of Paris was the most suitable oviposition substrate. Female L. shannoni adults laid no eggs on moist sand substrate. Sand fly adults held in groups of ten or more laid higher numbers of eggs than did individually maintained sand flies. Inclusion of the L. longipalpis oviposition hormone dodecanoic acid or the presence of previously laid eggs did not stimulate L. shannoni oviposition. The average L. shannoni egg, larval, and pupal duration were 9.3, 36.7, and 17.8 days, respectively. The addition of a 20% sugar solution improved adult female longevity. Females survived longer (14.8 days) than males (11.9 days). Lutzomyia shannoni was successfully colonized in the laboratory for up to four generations using this artificial membrane technique.     

The use of image analysis for spore counts of white-rot fungi

Summary An improved method has been developed for the determination of fungal spore counts using a haemocytometer. The method uses a personal computer with a frame grabber card and commercial imaging software. Semi-automation using an iterative programming technique was implemented to count the spores produced byPhanerochaete chrysosporium. Rapid and reliable counts were achieved using image processing.     

Adaptor Ligation-Based Polymerase Chain Reaction-Mediated Walking

An improved method of adaptor ligation PCR was developed for isolation of unknown sequences flanking a known DNA sequence. It was determined that the specificity of the adaptor ligation-based walking technique could be significantly enhanced by using uniquely blocked adaptors along with removal of unligated genomic DNA by exonuclease III digestion. This technique was utilized to isolate three novel promoter regions from three differentZea mays(maize) peroxidase genes. Sequences encoding a putative maize 6-phosphogluconate dehydrogenase gene were also isolated and confirmed by sequence analysis. The described improvements could be applied to other existing adaptor ligation-based PCR walking techniques.     

Improved technique for calculating X-ray scattering intensity of biopolymers in solution: Evaluation of the form,volume, and surface of a particle

An improved cube method has been developed for calculating the intensity of diffuse x-ray scattering of macromolecules in solution using a certain set of their atomic coordinates. The technique is based on the ideas of B. Lee and F. M. Richards [(1971) J. Mol. Biol. 55 , 374–400] and Richards [(1977) Annu. Rev. Biophys. Bioeng. 6 , 151–176] on the possibility of estimating the molecular and accessible surface of a particle by “rolling” a sphere, simulating a water molecule, on its molecular surface. It is shown that this technique is more advantageous than earlier versions of the cube methods. The improved technique for calculating scattering curves was utilized for several globular proteins, and for the first time, reliable scattering curves were obtained for protein-“bound” water complexes. In the case of globular proteins and tRNA, this technique has permitted a strict evaluation of their accessible surfaces, their volumes, and, apparently for the first time, their complete molecular surfaces.     

Ultrastructure of the epithelium from the ovary wall of Dendrochirus brachypterus (Pteroidae,Teleostei)

Summary The inner epithelium of the ovary wall in Dendrochirus brachypterus does not take part in egg production, as it does in other teleosts. The epithelium consists of columnar cells rich in mitochondria and secretory organelles. The distal ends of these cells hang free in the ovary lumen, separated from each other and densely covered by large microvilli. During reproduction the epithelium secretes large amounts of mucus that forms an envelope around the eggs produced from a spongy stroma of the ovary, and keeps the spawn afloat for 24 h.Thanks are due to Mrs. H. Segal and Mr. N. Sharon for technical assistance, and Mr. D. Fridman, for help during the collection of the fish. Also appreciation is given to the technicians of the Electron Microscopy Laboratory of Tel-Aviv University for help in preparation of electron and scanning microscope figures     

Automated extraction of bank angles from bird‐borne video footage using open‐source software

The use of miniaturized video cameras to study the at‐sea behavior of flying seabirds has increased in recent years. These cameras allow researchers to record several behaviors that were not previously possible to observe. However, video recorders produce large amounts of data and videos can often be time‐consuming to analyze. We present a new technique using open‐source software to extract bank angles from bird‐borne video footage. Bank angle is a key facet of dynamic soaring, which allows albatrosses and petrels to efficiently search vast areas of ocean for food. Miniaturized video cameras were deployed on 28 Wandering Albatrosses (Diomedea exulans) on Marion Island (one of the two Prince Edward Islands) from 2016 to 2018. The OpenCV library for the Python programming language was used to extract the angle of the horizon relative to the bird’s body (= bank angle) from footage when the birds were flying using a series of steps focused on edge detection. The extracted angles were not significantly different from angles measured manually by three independent observers, thus being a valid method to measure bank angles. Image quality, high wind speeds, and sunlight all influenced the accuracy of angle estimates, but post‐processing eliminated most of these errors. Birds flew most often with cross‐winds (58%) and tailwinds (39%), resulting in skewed distributions of bank angles when birds turned into the wind more often. Higher wind speeds resulted in extreme bank angles (maximum observed was 94°). We present a novel method for measuring postural data from seabirds that can be used to describe the fine‐scale movements of the dynamic‐soaring cycle. Birds appeared to alter their bank angle in response to varying wind conditions to counter wind drift associated with the prevailing westerly winds in the Southern Ocean. These data, in combination with fine‐scale positional data, may lead to new insights into dynamic‐soaring flight.     

Elimination of 2-deoxyribose interference in the thiobarbituric acid determination of N-acetylneuraminic acid in tumor cells by pH-dependent extraction with cyclohexanone

The widely used thiobarbituric acid technique for the quantitation of N-acetylneuraminic acid (NANA) was improved to eliminate the interference of the ubiquitous 2-deoxyribose. The 2-deoxyribose chromogen was completely removed by cyclohexanone extraction at pH 5.6–6.0. After readjusting the pH to 1.7–1.9, the chromogen representative of NANA was quantitatively extracted with cyclohexanone. All other aspects of the original technique (L. Warren 1959 J. Biol. Chem. 534, 1971–1975) remained unchanged. The technique has been applied to determine total as well as neuraminidase-susceptible NANA in the preparation of the immunogen (neuraminidase-treated myeloblasts) utilized to stimulate specific immunity in patients with myeloblastic leukemia and certain solid tumors; NANA levels significantly affect immunogenicity. Data obtained from a variety of tumors using pH-dependent extraction as compared to the thiobarbituric acid method, isoamyl alcohol extraction, and ion-exchange purification showed that 2-deoxyribose interference may cause as much a two- to threefold error in the quantitation of NANA.     

An improved night-lighting technique for the selective capture of sandgrouse and other steppe birds

The night-lighting technique is routinely used to catch birds. We improved this technique using a thermal infrared camera to catch steppe birds such as the sandgrouse. Target birds were located by an observer using the thermal camera, and approached and dazzled by another observer carrying a spotlight attached to a helmet and a hand-held net together with a playback noise to camouflage the footsteps of the observer. Using this improved technique, we caught on average 1.14 individuals per 3-h capture session (N = 81) and a total of 92 sandgrouse in 2007–2010 (86 pin-tailed sandgrouse Pterocles alchata and six black-bellied sandgrouse Pterocles orientalis). Capture rate and success were negatively influenced by moonlight (lowest during full moon nights). Our night-lighting technique is a highly selective and harmless method to capture sandgrouse and can be used for other small/medium-sized open-land birds or mammals.     

Kneading Technique for Preparation of Binary Solid Dispersion of Meloxicam with Poloxamer 188

The aim of the present study was to enhance the dissolution rate of meloxicam (MLX), a practically water-insoluble drug by preparation of solid dispersion using a hydrophilic polymer, poloxamer 188 (PXM). The kneading technique was used to prepare solid dispersions. A 3² full factorial design approach was used for optimization wherein the drug, polymer ratio (X ₁), and the kneading time (X ₂) were selected as independent variables and the dissolution efficiency at 60 min (%DE₆₀) and yield percent were selected as the dependent variable. Multiple linear regression analysis revealed that for obtaining higher dissolution of MLX from PXM solid dispersions, a high level of X ₁ and a high level of X ₂ were suitable. The use of a factorial design approach helped in optimization of the preparation and formulation of solid dispersion. The optimized formula was characterized by solubility studies, angle of repose, and contact angle; Fourier transform infrared spectroscopy, differential scanning calorimetry, x-ray diffraction studies, and scanning electron microscopy demonstrated that enhanced dissolution of MLX from solid dispersion might be due to a decrease in the crystallinity of MLX and PXM. Analysis of dissolution data of optimized formula indicated the best fitting with Korsemeyer–Peppas model and the drug release kinetics as Fickian diffusion. In conclusion, dissolution enhancement of MLX was obtained by preparing its solid dispersion with PXM using kneading technique.     

Protein structure calculation with data imputation: the use of substitute restraints

The amount of experimental restraints e.g., NOEs is often too small for calculating high quality three-dimensional structures by restrained molecular dynamics. Considering this as a typical missing value problem we propose here a model based data imputation technique that should lead to an improved estimation of the correct structure. The novel automated method implemented in AUREMOL makes a more efficient use of the experimental information to obtain NMR structures with higher accuracy. It creates a large set of substitute restraints that are used either alone or together with the experimental restraints. The new approach was successfully tested on three examples: firstly, the Ras-binding domain of Byr2 from Schizosaccharomyces pombe, the mutant HPr (H15A) from Staphylococcus aureus, and a X-ray structure of human ubiquitin. In all three examples, the quality of the resulting final bundles was improved considerably by the use of additional substitute restraints, as assessed quantitatively by the calculation of RMSD values to the “true” structure and NMR R-factors directly calculated from the original NOESY spectra or the published diffraction data.     

A standard karyogram of Petunia hybrida hort

An improved technique, viz., maceration with cellulase and pectinase, was applied in order to obtain chromosome preparations of botanical material suitable for fluorescence microscopy. The application of this method to Petunia hybrida allows the individual distinction of all chromosomes in mitosis. The following three criteria are required for this distinetion: the centromere index, the relative length, and the fluorescence intensity pattern after staining with Quinacrine. A standard karyogram is drawn up to render possible a comparison with other Petunia material.     

A versatile and general splitting technology for generating targeted YAC subclones

Yeast artificial chromosomes (YAC) splitting technology was developed as a means to subclone any desired region of eukaryotic chromosomes from one YAC into new YACs. In the present study, the conventional YAC splitting technology was improved by incorporating PCR-mediated chromosome splitting technique and by adding autonomously replicating sequence (ARS) to the system. To demonstrate the performance of the improved method, a 60-kb region from within a 590-kb YAC (clone CIC9e2 from Arabidopsis thaliana chromosome 5) that could not be subcloned using the original method was split to convert into a replicating YAC. Two template plasmids, pSK-KCA and pSKCLY, were used to generate two splitting fragments by PCR. Two splitting fragments consisted of telomeric (C₄A₂)₆ repeats, 400-bp target region, CEN4, H4ARS and Km^r (selective marker for plant transformants), or CgLEU2. These splitting fragments were introduced into Saccharomyces cerevisiae harboring the 100-kb split YAC generated by splitting of the 590-kb YAC and containing the 60-kb region. Among 12 Leu⁺ transformants, four exhibited the expected karyotype in which two newly split 40- and 60-kb chromosomes were generated. These results demonstrate that the improved method can convert a targeted region of a eukaryotic chromosome within a YAC into a replicating YAC.     

Towards Efficient Isolation of R Gene Orthologs from Multiple Genotypes: Optimization of Long Range-PCR

Resistance (R) genes and the proteins they encode are key components of the defense system of plants. The exploration of R gene diversity enables the study of R gene evolution and may facilitate the isolation of new and functional alleles. Most cloned R genes occur in clusters of related sequences. Thus, the development of a tool for reliable recovery of orthologous R gene sequences to the exclusion of paralogous sequences will facilitate R gene diversity analysis. The late blight resistance gene RB is a single functional locus embedded within a cluster of related sequences. Previously, the functional RB allele was cloned from wild potato using a Long Range-PCR (LR-PCR) technique, suggesting this method may be a promising tool for recovery of R gene orthologs in other genotypes. Using the RB gene as a model, we explored the limitations and improved three technical aspects of LR-PCR for multi-genotype applications. We present improved primers for the recovery of the RB locus and have identified efficient DNA extraction procedures and reliable amplification systems. We document that consensus sequences built from three independently generated LR-PCR clones can be up to 100% accurate. Our results show encouraging advances toward successful application of LR-PCR for isolating alleles from orthologous R gene loci.     

Measuring and evaluating morphological asymmetry in fish: distinct lateral dimorphism in the jaws of scale‐eating cichlids

The left–right asymmetry of scale‐eating Tanganyikan cichlids is described as a unilateral topographical shift of the quadratomandibular joints. This morphological laterality has a genetic basis and has therefore been used as a model for studying negative frequency‐dependent selection and the resulting oscillation in frequencies of two genotypes, lefty and righty, in a population. This study aims were to confirm this laterality in Perissodus microlepis Boulenger and P. straeleni (Poll) and evaluate an appropriate method for measuring and testing the asymmetry. Left–right differences in the height of the mandible posterior ends (HMPE) and the angle between the neurocranium and vertebrae of P. microlepis and P. straeleni were measured on skeletal specimens. Snout‐bending angle was also measured using a dorsal image of the same individuals following a previous method. To define which distribution model, fluctuating asymmetry (FA), directional asymmetry (DA), or antisymmetry (AS), best fit to the lateral asymmetry of the traits, we provided an R package, IASD. As a result, HMPE and neurocranium–vertebrae angle of both species were best fitted to AS, suggesting that P. microlepis and P. straeleni showed a distinct dimorphism in these traits, although snout‐bending angle of P. microlepis was best fitted to FA. Measurement error was low for HMPE comparing the snout‐bending angle in P. microlepis, indicating that measuring HMPE is a more accurate method. The scale‐eating tribe Perissodini showed distinct antisymmetry in the jaw skeleton and neurocranium–vertebrae angle, and this laterality remains a valid marker for further evolutionary studies.     

Optimization of interatomic angle geometry in torsion angle space

Summary Interatomic angle constraints are usually specified as distance constraints in torsion angle distance geometry. Such an approach is inaccurate and often inadequate. We provide a direct definition of the inter-atomic angle constraint term, which can be incorporated in the target function. The first derivative of this term with respect to the torsion angle has been described for all possible cases. This feature has been implemented in the nucleic acid distance geometry program TANDY [Ajay Kumar et al. (1991)J. Biomol NMR,1, 363–378], and has been tested on base pairing in the DNA fragment, d(AT)₂. The results clearly indicate the need and adequacy of such angle constraints. Other applications that would also benefit from this technique have been identified.     

Extra thin alginate film: an efficient technique for protoplast culture

Summary. This paper reports an efficient protoplast culture technique, the “extra thin alginate film” technique. The development of this improved method of protoplast culture was an outcome of an assessment of the efficiency and shortcomings of various protoplast culture techniques. The efficiency of this technique was evaluated with two model plant systems, viz., Nicotiana tabacum and Lotus corniculatus, and a comparison was made with the “thin alginate layer” technique, another efficient protoplast culture system. Results indicate that the culture technique with extra thin alginate film is as efficient as the technique with thin alginate layer, with many additional advantages. The present innovation overcomes most of the limitations of protoplast culture techniques described so far and can now be applied to a wide variety of crops to check its general applicability. Correspondence and reprints: Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar 143 005, India.