PREDICTION OF FAT-FREE MASS BY BIOELECTRICAL IMPEDANCE ANALYSIS IN OLDER ADULTS FROM DEVELOPING COUNTRIES: A CROSS-VALIDATION STUDY USING THE DEUTERIUM DILUTION METHOD

Objective: Several limitations of published bioelectrical impedance analysis (BIA) equations have been reported. The aims were to develop in a multiethnic, elderly population a new prediction equation and cross-validate it along with some published BIA equations for estimating fat-free mass using deuterium oxide dilution as the reference method. Design and setting: Cross-sectional study of elderly from five developing countries. Methods: Total body water (TBW) measured by deuterium dilution was used to determine fat-free mass (FFM) in 383 subjects. Anthropometric and BIA variables were also measured. Only 377 subjects were included for the analysis, randomly divided into development and cross-validation groups after stratified by gender. Stepwise model selection was used to generate the model and Bland Altman analysis was used to test agreement. Results: FFM = 2.95 - 3.89 (Gender) + 0.514 (Ht(2)/Z) + 0.090 (Waist) + 0.156 (Body weight). The model fit parameters were an R(2), total F-Ratio, and the SEE of 0.88, 314.3, and 3.3, respectively. None of the published BIA equations met the criteria for agreement. The new BIA equation underestimated FFM by just 0.3 kg in the cross-validation sample. The mean of the difference between FFM by TBW and the new BIA equation were not significantly different; 95% of the differences were between the limits of agreement of -6.3 to 6.9 kg of FFM. There was no significant association between the mean of the differences and their averages (r = 0.008 and p = 0.2). Conclusions: This new BIA equation offers a valid option compared with some of the current published BIA equations to estimate FFM in elderly subjects from five developing countries.

Evaluation of bioactivities of a propolis sample (Gerês) of Portuguese origin

Dissertação de mestrado em Genética Molecular

Validity of impedance-based equations for the prediction of total body water as measured by deuterium dilution in African women.

BACKGROUND: Little information is available on the validity of simple and indirect body-composition methods in non-Western populations. Equations for predicting body composition are population-specific, and body composition differs between blacks and whites. OBJECTIVE: We tested the hypothesis that the validity of equations for predicting total body water (TBW) from bioelectrical impedance analysis measurements is likely to depend on the racial background of the group from which the equations were derived. DESIGN: The hypothesis was tested by comparing, in 36 African women, TBW values measured by deuterium dilution with those predicted by 23 equations developed in white, African American, or African subjects. These cross-validations in our African sample were also compared, whenever possible, with results from other studies in black subjects. RESULTS: Errors in predicting TBW showed acceptable values (1.3-1.9 kg) in all cases, whereas a large range of bias (0.2-6.1 kg) was observed independently of the ethnic origin of the sample from which the equations were derived. Three equations (2 from whites and 1 from blacks) showed nonsignificant bias and could be used in Africans. In all other cases, we observed either an overestimation or underestimation of TBW with variable bias values, regardless of racial background, yielding no clear trend for validity as a function of ethnic origin. CONCLUSIONS: The findings of this cross-validation study emphasize the need for further fundamental research to explore the causes of the poor validity of TBW prediction equations across populations rather than the need to develop new prediction equations for use in Africa.

Random measurement error and regression dilution bias.

Summary points: - The bias introduced by random measurement error will be different depending on whether the error is in an exposure variable (risk factor) or outcome variable (disease) - Random measurement error in an exposure variable will bias the estimates of regression slope coefficients towards the null - Random measurement error in an outcome variable will instead increase the standard error of the estimates and widen the corresponding confidence intervals, making results less likely to be statistically significant - Increasing sample size will help minimise the impact of measurement error in an outcome variable but will only make estimates more precisely wrong when the error is in an exposure variable

Synthesis of isotope-labelled methoxypyrazine compounds as internal standards and quantitative determination of aroma methoxypyrazines in water and wines by solid-phase extraction with isotope dilution-GC-MS /

An efficient way of synthesizing the deuterium labelled analogues of three methoxypyrazine compounds: 2-d3-methoxy-3-isopropylpyrazine, 2-d3-methoxy-3- isobutylpyrazine, and 2-d3-methoxy-3-secbutylpyrazine, has been developed. To confirm that the deuterium labels had been incorporated into the expected positions in the molecules synthesized, the relevant characterization by NMR, HRMS and GC/MS analysis was conducted. Another part of this work involved quantitative determination of methoxypyrazines in water and wines. Solid-phase extraction (SPE) proved to be a suitable means for the sample separation and concentration prior to GC/MS analysis.Such factors as the presence of ethanol, salt, and acid have been investigated which can influence the recovery by SPE for the pyrazines from the water matrix. Significantly, in this work comparatively simple fractional distillation was attempted to replace the conventional steam distillation for pre-concentrating a sample with a relatively large volume prior to SPE. Finally, a real wine sample spiked with the relevant isotope-labelled methoxypyrazines was quantitatively analyzed, revealing that the wine with 10 beetles per litre contained 138 ppt of 2-methoxy-3-isopropylpyrazine. Interestingly, we have also found that 2-methoxy-3-secbutylpyrazine exhibits an extremely low detection limit in GC/MS analysis compared with the detection limit of the other two methoxypyrazines: 2- methoxy-3-isopropylpyrazine and 2-methoxy-3-isobutylpyrazine.

Estimating numbers of infectious units from serial dilution assays

The paper concerns the design and analysis of serial dilution assays to estimate the infectivity of a sample of tissue when it is assumed that the sample contains a finite number of indivisible infectious units such that a subsample will be infectious if it contains one or more of these units. The aim of the study is to estimate the number of infectious units in the original sample. The standard approach to the analysis of data from such a study is based on the assumption of independence of aliquots both at the same dilution level and at different dilution levels, so that the numbers of infectious units in the aliquots follow independent Poisson distributions. An alternative approach is based on calculation of the expected value of the total number of samples tested that are not infectious. We derive the likelihood for the data on the basis of the discrete number of infectious units, enabling calculation of the maximum likelihood estimate and likelihood-based confidence intervals. We use the exact probabilities that are obtained to compare the maximum likelihood estimate with those given by the other methods in terms of bias and standard error and to compare the coverage of the confidence intervals. We show that the methods have very similar properties and conclude that for practical use the method that is based on the Poisson assumption is to be recommended, since it can be implemented by using standard statistical software. Finally we consider the design of serial dilution assays, concluding that it is important that neither the dilution factor nor the number of samples that remain untested should be too large.

Use of FIA systems for on-line dilution in multielement determination by ICP-MS

A flow-injection (FI) system to match concentrations was used as an auto-diluter in multielement determination by inductively coupled plasma-mass spectrometry (ICP-MS). The flow system comprised loop-based injection or a timed valve that introduced a variable sample volume info a spray chamber through a standard Meinhard nebulizer of an ICP-MS. Routinely analyzed samples such as water, plant, and steel were selected. The accuracy of multielement determination was checked against water standard reference material from the National Institute of Standards and Technology (1643d), plant standard reference material from the National Bureau of Standards (1572 citrus leaves), and steel standard reference material from the National Bureau of Standards (AISI 4340). The measuring system was calibrated with a multielement solution, yielding a linear plot with good precision [relative standard deviation (RSD) < 3%, n = 12]. The results were in agreement at a 95% confidence level with the certified values for the reference materials and also with those obtained by continuous aspiration and by (FI) with a discrete volume. (C) 1999 John Wiley & Sons, Inc.

Determination of trace elements in serum samples by isotope dilution inductively coupled plasma mass spectrometry using on-line dialysis

An on-line dialysis flow system coupled to inductively coupled plasma mass spectrometry to determine trace elements in serum samples by isotope dilution is presented. Isotope dilution was performed on samples incubated with enriched Cu-65, Zn-66, Se-77 and Pb-206 for 24 h at 36degreesC prior to dialysis to quantified total element concentrations. The sample and acceptor solutions flowed through the dialysis unit with cellophane membrane placed in between the compartments. The serum sample (1 mL) was left to recycle in a closed path while the acceptor solution was continuously pumped along the dialyzer channel and through a cationic AG50W X-8 resin column. After 10 min, around 70% of Na, K and Cl migrate from the sample. Three replicate injections of 0.1 mL were performed for the clean sample after each separation step. The on-line coupling of the dialyzer to ICP-MS allowed isotope dilution for total element determination either in the cleaned sample or by eluting the cations retained in the resin to be carried out. Results demonstrated no matrix effects from alkaline elements or spectral interference from ArNa+ on Cu-63, ArCl+ on Se-77 and (SO2+)-S-34 on Zn-66. The precision of isotope ratio measurements for Cu and Zn was around 1% and for Se and Pb was around 2.5%. The values found for the reference serum sample IMEP-17 were in good agreement with the certified values for Cu, Zn and Se.

Dilution of candidates: the case of iron-related genes in restless legs syndrome

Restless legs syndrome (RLS) is a common multifactorial disease. Some genetic risk factors have been identified. RLS susceptibility also has been related to iron. We therefore asked whether known iron-related genes are candidates for association with RLS and, vice versa, whether known RLS-associated loci influence iron parameters in serum. RLS/control samples (n = 954/1814 in the discovery step, 735/736 in replication 1, and 736/735 in replication 2) were tested for association with SNPs located within 4 Mb intervals surrounding each gene from a list of 111 iron-related genes using a discovery threshold of P = 5 × 10(-4). Two population cohorts (KORA F3 and F4 with together n = 3447) were tested for association of six known RLS loci with iron, ferritin, transferrin, transferrin-saturation, and soluble transferrin receptor. Results were negative. None of the candidate SNPs at the iron-related gene loci was confirmed significantly. An intronic SNP, rs2576036, of KATNAL2 at 18q21.1 was significant in the first (P = 0.00085) but not in the second replication step (joint nominal P-value = 0.044). Especially, rs1800652 (C282Y) in the HFE gene did not associate with RLS. Moreover, SNPs at the known RLS loci did not significantly affect serum iron parameters in the KORA cohorts. In conclusion, the correlation between RLS and iron parameters in serum may be weaker than assumed. Moreover, in a general power analysis, we show that genetic effects are diluted if they are transmitted via an intermediate trait to an end-phenotype. Sample size formulas are provided for small effect sizes.

Nd and Sr isotopic compositions and isotope dilution trace element concentrations from 7 sediment cores along the southeast Greenland margin

The Nd and Sr isotopic compositions of Quaternary glacial and glacimarine siliciclastic sediments deposited along the margin of southeast Greenland were determined to assess the roles of the Greenland, Iceland, and more distal ice sheets in delivering detritus to this portion of the northern North Atlantic. The isotopic compositions of detritus generated by portions of the southern Greenland Ice Sheet were defined through measurements of till and trough mouth fan sediments. Massive diamicts from the Scoresby Sund trough mouth fan show a restricted range of e-Nd (-11.8 to -16.6) and 87Sr/86Sr (0.7192-0.7246) consistent with their derivation from mixtures of sediments derived from Paleoproterozoic and/or Caledonian basement and Tertiary Greenland basalts. Further south at Kangerlussuaq, till isotopic compositions covary with the underlying basement type, with low e-Nd values in the inner fiord (-18.1) reflecting the erosion of the local Precambrian gneisses, but with higher e-Nd values (-2.3 to 2.5) found where the trough crosses East Greenland Tertiary basalts. Fine-grained (< 63 µm) sediments deposited along the southeast Greenland margin also show regular spatial isotopic variations. Ambient sediments and ice-rafted detritus in the southern Irminger Basin trend towards low e-Nd values (to ~ -28) and 87Sr/86Sr ratios (~ 0.711 to ~ 0.715) and are likely derived from proximal Archean gneisses of SE Greenland. Further north in the northern Irminger and Blosseville Basins, sediments trend toward much higher e-Nd (> -4) and low 87Sr/86Sr (< 0.709) reflecting a component derived from the local Iceland volcanic rocks and/or the East Greenland Tertiary basalts. In all three regions, the locally-derived detritus is intermixed with sediment with an intermediate e-Nd value (~ -10) and 87Sr/86Sr (~ 0.718) that was likely delivered by icebergs emanating from the Eurasian Ice Sheets and not from eastern Greenland. Deposition of glacial sediments from both proximal and distal (Eurasian) sources occurred adjacent to SE Greenland throughout the past 50 Ka, with periodic increases in IRD deposition at various times including those of Heinrich events 1, 2 and 4. These results suggest that at least the southern portions of the Greenland Ice Sheet experienced periodic instabilities during the Last Glacial period.