Segmental body composition assessment for obese Japanese adults by single-frequency bioelectrical impedance analysis with 8-point contact electrodes

This study aimed to determine the accuracy of segmental body composition variables estimated by single-frequency BIA with 8-point contact electrodes (SF-BIA8), compared with dual-energy X-ray absorptiometry (DXA). Subjects were 72 obese Japanese adults (43 males and 29 females) aged 30 to 66 years. Segmental body composition variables (fat free mass: FFM, fat mass: FM, and percent fat mass: %FAT) were measured by these techniques. The correlations between impedance values and FFM measured by DXA were calculated. To examine the consistency in predicted values (SF-BIA8) with the reference (DXA), significant mean differences were tested by t-test and the degree of the difference was assessed by effect size. Correlations between the reference and predicted values were calculated. Additionally, the standard error of estimation (SEE) when estimating the reference from the predictor and the relative value of the SEE to the mean value of the DXA measurement (%SEE) were calculated. Systematic error was examined by Bland-Altman plots. High correlations were found between impedance and FFM measured by SF-BIA8. FFM in the extremities showed high correlations with the reference values, but systematic error was found. SF-BIA8 tended to overestimate FFM in the trunk. The consistencies in %FAT and FM with the reference value are inferior to those for FFM, and SEE values in %FAT and FM were greater than those for FFM. The accuracy of the estimated values in the trunk (FFM, %FAT, and FM) are inferior to those of the total body and extremities.     

Gender differences in body-sway factors of center of foot pressure in a static upright posture and under the influence of alcohol intake

This study aimed to examine gender differences in 4 body-sway factors of the center of foot pressure (CFP) during a static upright posture and the influence of alcohol intake on them. Four body-sway factors were interpreted in previous studies using factor analysis (the principal factor method and oblique solution by promax-rotation) on 220 healthy young males and females as follows; unit time sway, front-back sway, left-right sway and high frequency band power. The CFP measurement for 1 min was carried out twice with 1 min rest. The measurements of blood pressure, heart rate, whole body reaction time, standing on one leg with eyes closed, and CFP were carried out before and after the alcohol intake using 11 healthy young males and females. The measurement device used was an Anima's stabilometer G5500. The data sampling frequency was 20 Hz. Reliability of 4 body-sway factors was very high. Significant gender differences were found in the left-right sway and the high frequency band power factors, but the influence on body-sway is, as a whole, can be disregarded. These four sway factors can determine the influence of alcohol intake as efficient as 32 sway parameters.     

Percentage of total body fat as estimated by three automatic bioelectrical impedance analyzers

The present study aimed to compare the accuracy of estimating the percentage of total body fat (%TBF) among three bioelectrical impedance analysis (BIA) devices: a single-frequency BIA with four tactile electrodes (SF-BIA4), a single-frequency BIA with eight tactile electrodes (SF-BIA8) and a multi-frequency BIA with eight tactile electrodes (MF-BIA8). Dual-energy x-ray absorptiometry (DXA) and hydrostatic weighing (HW) were used as references for the measured values. Forty-five healthy college student volunteers (21 males: 172.9 +/- 5.5 cm and 65.8 +/- 9.1 kg and 24 females: 160.7 +/- 6.6 cm, 52.6 +/- 6.2 kg) were the subjects. Correlation coefficients between the BIA measurements and the references were calculated. The standard error of estimation (SEE) was calculated by regression analysis when estimating the reference measures (DXA and HW) from the predictor (SF-BIA4, SF-BIA8 and MF-BIA8). The differences in %TBF between the reference and the predictor, calculated by the reference minus the predictor, were plotted against the %TBF measured by the references. The MF-BIA 8 here showed the highest correspondence to the reference and the least estimation error compared with the other BIA methods. It is considered that there is a limit to directly estimate FFM from a regression equation using impedance, weight, height and age as independent variables, and that %TBF can be more accurately estimated by measuring segmental impedances using eight electrodes and multi-frequency electric currents and then estimating total body water from these impedances.     

Examination of the factor structure of center of foot pressure movement and cross-validity

This study aimed to determine the factor structure of the center of foot pressure (CFP) movement during static upright posture, and to objectively categorize and summarize parameters to evaluate CFP movement. The subjects were 220 healthy young males and females. The measurement of CFP was carried out 3 times with 1 min rest and the mean of trials 2 and 3 was used for the analysis. The measurement device was an Anima's stabilometer G5500. The data sampling frequency was 20 Hz. Thirty-four parameters with high reliability were selected from the following 6 domains except for the center position which is a fundamental attribute: distance, distribution of amplitude, area, velocity, power spectrum, and body sway vector. Factor analysis (principal factor method and promax rotation) was applied to a correlation matrix consisting of 32 parameters. Four factors abstracted were interpreted as follows; unit time sway, front and back sway, left and right sway and high frequency band of power spectrum. The reliability coefficient (ICC=0.89-0.95) and the congruence coefficient (phi=0.80-0.97) between factors abstracted from the original and the cross-validity groups were very high. It was considered that the CFP movement consists of the above 4 factors that evaluate the amount of body sway and can be synthetically evaluated by them.     

Gene cloning and function analysis of replication factor C from Thermococcus kodakaraensis KOD1

Replication factor C (RFC) catalyzes the assembly of circular proliferating cell nuclear antigen (PCNA) clamps around primed DNA, enabling processive synthesis by DNA polymerase. The RFC-like genes, arranged in tandem in the Thermococcus kodakaraensis KOD1 genome, were cloned individually and co-expressed in Escherichia coli cells. T. kodakaraensis KOD1 RFC homologue (Tk-RFC) consists of the small subunit (Tk-RFCS: MW=37.2 kDa) and the large subunit (Tk-RFCL: MW=57.2 kDa). The DNA elongation rate of the family B DNA polymerase from T. kodakaraensis KOD1 (KOD DNA polymerase), which has the highest elongation rate in all thermostable DNA polymerases, was increased about 1.7 times, when T. kodakaraensis KOD1 PCNA (Tk-PCNA) and the Tk-RFC at the equal molar ratio of KOD DNA polymerase were reacted with primed DNA.     

hHGF overexpression in myoblast sheets enhances their angiogenic potential in rat chronic heart failure

After severe myocardial infarction (MI), heart failure results from ischemia, fibrosis, and remodeling. A promising therapy to enhance cardiac function and induce therapeutic angiogenesis via a paracrine mechanism in MI is myoblast sheet transplantation. We hypothesized that in a rat model of MI-induced chronic heart failure, this therapy could be further improved by overexpression of the antiapoptotic, antifibrotic, and proangiogenic hepatocyte growth factor (HGF) in the myoblast sheets. We studied the ability of wild type (L6-WT) and human HGF-expressing (L6-HGF) L6 myoblast sheet-derived paracrine factors to stimulate cardiomyocyte, endothelial cell, or smooth muscle cell migration in culture. Further, we studied the autocrine effect of hHGF-expression on myoblast gene expression profiles by use of microarray analysis. We induced MI in Wistar rats by left anterior descending coronary artery (LAD) ligation and allowed heart failure to develop for 4 weeks. Thereafter, we administered L6-WT (n = 15) or L6-HGF (n = 16) myoblast sheet therapy. Control rats (n = 13) underwent LAD ligation and rethoracotomy without therapy, and five rats underwent a sham operation in both surgeries. We evaluated cardiac function with echocardiography at 2 and 4 weeks after therapy, and analyzed cardiac angiogenesis and left ventricular architecture from histological sections at 4 weeks. Paracrine mediators from L6-HGF myoblast sheets effectively induced migration of cardiac endothelial and smooth muscle cells but not cardiomyocytes. Microarray data revealed that hHGF-expression modulated myoblast gene expression. In vivo, L6-HGF sheet therapy effectively stimulated angiogenesis in the infarcted and non-infarcted areas. Both L6-WT and L6-HGF therapies enhanced cardiac function and inhibited remodeling in a similar fashion. In conclusion, L6-HGF therapy effectively induced angiogenesis in the chronically failing heart. Cardiac function, however, was not further enhanced by hHGF expression.     

Structural mechanism for coordination of proofreading and polymerase activities in archaeal DNA polymerases

A novel mechanism for controlling the proofreading and polymerase activities of archaeal DNA polymerases was studied. The 3'-5'exonuclease (proofreading) activity and PCR performance of the family B DNA polymerase from Thermococcus kodakaraensis KOD1 (previously Pyrococcus kodakaraensis KOD1) were altered efficiently by mutation of a "unique loop" in the exonuclease domain. Interestingly, eight different H147 mutants showed considerable variations in respect to their 3'-5'exonuclease activity, from 9% to 276%, as against that of the wild-type (WT) enzyme. We determined the 2.75A crystal structure of the H147E mutant of KOD DNA polymerase that shows 30% of the 3'-5'exonuclease activity, excellent PCR performance and WT-like fidelity. The structural data indicate that the properties of the H147E mutant were altered by a conformational change of the Editing-cleft caused by an interaction between the unique loop and the Thumb domain. Our data suggest that electrostatic and hydrophobic interactions between the unique loop of the exonuclease domain and the tip of the Thumb domain are essential for determining the properties of these DNA polymerases.